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/*===========================================================================
*
* PUBLIC DOMAIN NOTICE
* National Center for Biotechnology Information
*
* This software/database is a "United States Government Work" under the
* terms of the United States Copyright Act. It was written as part of
* the author's official duties as a United States Government employee and
* thus cannot be copyrighted. This software/database is freely available
* to the public for use. The National Library of Medicine and the U.S.
* Government have not placed any restriction on its use or reproduction.
*
* Although all reasonable efforts have been taken to ensure the accuracy
* and reliability of the software and data, the NLM and the U.S.
* Government do not and cannot warrant the performance or results that
* may be obtained by using this software or data. The NLM and the U.S.
* Government disclaim all warranties, express or implied, including
* warranties of performance, merchantability or fitness for any particular
* purpose.
*
* Please cite the author in any work or product based on this material.
*
* ===========================================================================
*
*/
/* #include "bam-load.vers.h" */
#ifdef __cplusplus
extern "C" {
#endif
#include <klib/callconv.h>
#include <klib/data-buffer.h>
#include <klib/text.h>
#include <klib/log.h>
#include <klib/out.h>
#include <klib/status.h>
#include <klib/rc.h>
#include <klib/sort.h>
#include <klib/printf.h>
#include <kfs/directory.h>
#include <kfs/file.h>
#include <kdb/btree.h>
#include <kdb/manager.h>
#include <kdb/database.h>
#include <kdb/table.h>
#include <kdb/meta.h>
#include <vdb/manager.h>
#include <vdb/schema.h>
#include <vdb/database.h>
#include <vdb/table.h>
#include <vdb/cursor.h>
#include <vdb/vdb-priv.h>
#include <insdc/insdc.h>
#include <insdc/sra.h>
#include <align/dna-reverse-cmpl.h>
#include <align/align.h>
#include <kapp/main.h>
#include <kapp/args.h>
#include <kapp/log-xml.h>
#include <kproc/queue.h>
#include <kproc/thread.h>
#include <kproc/timeout.h>
#include <os-native.h>
#include <loader/loader-file.h>
#include <loader/loader-meta.h>
#include <loader/progressbar.h>
#include <sysalloc.h>
#include <atomic32.h>
#include <unistd.h>
#include <fcntl.h>
#include <sys/mman.h>
#include <stdlib.h>
#include <stdio.h>
#include <string.h>
#include <ctype.h>
#include <assert.h>
#include <limits.h>
#include <time.h>
#include <zlib.h>
#include "bam.h"
#include "Globals.h"
#include "sequence-writer.h"
#include "reference-writer.h"
#include "alignment-writer.h"
#include "mem-bank.h"
#include "low-match-count.h"
#include "bam-alignment.h"
#ifdef __cplusplus
}
#endif
#include <spdlog/fmt/fmt.h>
#include <spdlog/spdlog.h>
#include <spdlog/sinks/stdout_sinks.h>
#include <spdlog/sinks/null_sink.h>
#include <spdlog/stopwatch.h>
#include <fstream>
#include "data_frame.hpp"
#include <taskflow/taskflow.hpp>
#include <taskflow/algorithm/sort.hpp>
#include <bm/bmsparsevec_algo.h>
#include <bm/bmtimer.h>
#include "hashing.hpp"
#ifdef __linux__
#include <sys/resource.h>
#endif
#include <tsl/array_map.h>
#include "spot_assembly.hpp"
#define NEW_QUEUE
#if defined(NEW_QUEUE)
#include "rwqueue/readerwriterqueue.h"
#endif
//#define HAS_CTX_VALUE 1
//#define NO_METADATA 1
using namespace std;
using namespace moodycamel;
#define NUM_ID_SPACES (256u)
#if defined(HAS_CTX_VALUE)
static constexpr unsigned MAX_GROUP_BITS = 32;
#else
static constexpr unsigned MAX_GROUP_BITS = 24;
#endif
static constexpr unsigned GROUPID_SHIFT = (64 - MAX_GROUP_BITS);
static constexpr uint64_t KEYID_MASK = ~(~(uint64_t)0 << GROUPID_SHIFT);
static constexpr unsigned MAX_GROUPS_ALLOWED = NUM_ID_SPACES;//(1u << MAX_GROUP_BITS);
#define MMA_NUM_CHUNKS_BITS (20u)
#define MMA_NUM_SUBCHUNKS_BITS ((32u)-(MMA_NUM_CHUNKS_BITS))
#define MMA_SUBCHUNK_SIZE (1u << MMA_NUM_CHUNKS_BITS)
#define MMA_SUBCHUNK_COUNT (1u << MMA_NUM_SUBCHUNKS_BITS)
/**
* Returns the current resident memory use measured in bytes
*/
size_t getCurrentRSS( )
{
long rss = 0L;
FILE* fp = NULL;
if ( (fp = fopen( "/proc/self/statm", "r" )) == NULL )
return (size_t)0L; /* Can't open? */
if ( fscanf( fp, "%*s%ld", &rss ) != 1 )
{
fclose( fp );
return (size_t)0L; /* Can't read? */
}
fclose( fp );
return (size_t)rss * (size_t)sysconf( _SC_PAGESIZE);
}
#if defined(HAS_CTX_VALUE)
typedef struct {
int fd;
size_t elemSize;
off_t fsize;
uint8_t *current;
struct mma_map_s {
struct mma_submap_s {
uint8_t *base;
} submap[MMA_SUBCHUNK_COUNT];
} map[NUM_ID_SPACES];
} MMArray;
typedef struct {
uint32_t primaryId[2];
uint32_t spotId;
uint32_t fragmentId;
uint8_t fragment_len[2]; /*** lowest byte of fragment length to prevent different sizes of primary and secondary alignments **/
uint8_t platform;
uint8_t pId_ext[2];
uint8_t spotId_ext;
uint8_t alignmentCount[2]; /* 0..254; 254: saturated max; 255: special meaning "too many" */
uint8_t unmated: 1,
pcr_dup: 1,
unaligned_1: 1,
unaligned_2: 1,
hardclipped: 1,
primary_is_set: 1;
} ctx_value_t;
#define CTX_VALUE_SET_P_ID(O,N,V) do { int64_t tv = (V); (O).primaryId[N] = (uint32_t)tv; (O).pId_ext[N] = tv >> 32; } while(0);
#define CTX_VALUE_GET_P_ID(O,N) ((((int64_t)((O).pId_ext[N])) << 32) | (O).primaryId[N])
#define CTX_VALUE_SET_S_ID(O,V) do { int64_t tv = (V); (O).spotId = (uint32_t)tv; (O).spotId_ext = tv >> 32; } while(0);
#define CTX_VALUE_GET_S_ID(O) ((((int64_t)(O).spotId_ext) << 32) | (O).spotId)
#endif
typedef struct
{
vector<uint32_t> values;
vector<uint8_t> ext;
uint64_t get(size_t index) const {
uint64_t v = ext[index];
v <<= 32;
v |= values[index];
return v;
}
} u40_t;
typedef struct FragmentInfo {
uint64_t ti;
uint32_t readlen;
uint8_t aligned;
uint8_t is_bad;
uint8_t orientation;
uint8_t readNo;
uint8_t sglen;
uint8_t lglen;
uint8_t cskey;
} FragmentInfo;
/**
* @brief Data returned by bam_read threads
*
*/
typedef struct
{
BAM_Alignment* alignment{nullptr}; ///< BAM Alignment
metadata_t* metadata{nullptr}; ///< Pointer to metadata
uint32_t row_id{0}; ///< Corresponding metadata row
} queue_rec_t;
typedef struct context_t {
array<const KLoadProgressbar*, 4> progress = {nullptr, nullptr, nullptr, nullptr};
MemBank *frags = nullptr;
uint64_t spotId = 0;
uint64_t primaryId = 0;
uint64_t secondId = 0;
uint64_t alignCount = 0;
unsigned pass;
bool isColorSpace;
BAM_FilePosition m_fileOffset = 0; ///< Position in the current BAM file
uint64_t m_inputSize = 0; ///< Total size in bytes of all input files (can be 0 for stdin inputs)
uint64_t m_processedSize = 0; ///< Number of already processed bytes
size_t m_estimatedBatchSize = 0; ///< Estimated size of the search batch
bool m_calcBatchSize = true; ///< Flag to indicate whether the batch needs to be calculated
unique_ptr<tf::Executor> m_executor; ///< Taskflow executor
#if defined(HAS_CTX_VALUE)
MMArray *id2value;
#endif
bool m_isSingleGroup{false}; ///< All reads belong to a single group (case when the number of groups in the header exceeds the allowed number)
unsigned m_emptyGroupIndex = -1; /**< When m_isSingleGroup is set, the index indicates that single group index,
important when there are multiple inputs and one of them is determined to be singleGroup one*/
using group_name_t = char;
using group_index_t = uint32_t;
tsl::array_map<group_name_t, group_index_t> m_group_map; ///< group name to group index
vector<unique_ptr<spot_assembly>> m_read_groups; ///< list of read groups
shared_ptr<spot_name_filter> m_key_filter; ///< Bloom filter (all spot names in scope)
vector<u40_t> m_spot_id_buffer; ///< Temporary buffer for spot name extraction
// reset everything but spotId and spot assembly related fields
void reset_for_remap() {
for (const auto& p : progress) {
if (p != nullptr)
KLoadProgressbar_Release(p, true);
}
frags = nullptr;
primaryId = 0;
secondId = 0;
alignCount = 0;
pass = 0;
isColorSpace = false;
}
/**
* @brief Set bloom filter based on the estimated number of spots
* >9e9 spots - sha256
* >2e9 - sha242
* >1e9 - sha1
* otherwise default: fnv + murmur
*
* @param num_spots
*/
void set_key_filter(size_t num_spots) {
assert(num_spots > 0);
static bool is_set = false;
if (is_set)
return;
if (num_spots > 3e9 && dynamic_cast<sha256_filter*>(m_key_filter.get()) == 0) {
spdlog::stopwatch sw;
m_key_filter.reset(new sha256_filter);
for(auto& gr : m_read_groups) {
gr->visit_spots([this](const char* spot_name) {
m_key_filter->seen_before(spot_name, strlen(spot_name));
});
}
spdlog::info("SHA256 bloom filter rebuilt in {:.3}", sw);
} else if (num_spots > 2e9 && dynamic_cast<sha224_filter*>(m_key_filter.get()) == 0) {
spdlog::stopwatch sw;
m_key_filter.reset(new sha224_filter);
for(auto& gr : m_read_groups) {
gr->visit_spots([this](const char* spot_name) {
m_key_filter->seen_before(spot_name, strlen(spot_name));
});
}
spdlog::info("SHA224 bloom filter rebuilt in {:.3}", sw);
} else if (num_spots > 1e9 && dynamic_cast<sha1_filter*>(m_key_filter.get()) == 0) {
spdlog::stopwatch sw;
m_key_filter.reset(new sha1_filter);
for(auto& gr : m_read_groups) {
gr->visit_spots([this](const char* spot_name) {
m_key_filter->seen_before(spot_name, strlen(spot_name));
});
}
spdlog::info("SHA1 bloom filter rebuilt in {:.3}", sw);
}
is_set = true;
}
spot_assembly& add_read_group() {
m_read_groups.emplace_back(make_unique<spot_assembly>(*m_executor, m_key_filter, m_read_groups.size(), G.searchBatchSize));
return *m_read_groups.back().get();
}
/**
* @brief Release memory spot assembly memory (volume's data, index. scanner). It doesn't touch metadata
*
*/
void release_search_memory() {
m_group_map.clear();
m_group_map.shrink_to_fit();
m_key_filter.reset();
for (auto&& s : m_read_groups) {
s->release_search_memory();
}
}
/*
#if defined(HAS_CTX_VALUE)
template<typename F>
void visit_keyId(F&& f) {
unsigned group_id = 0;
for (auto& gr : m_read_groups) {
gr->visit_keyId(f, group_id, GROUPID_SHIFT, metadata_t::e_fragmentId);
++group_id;
}
}
#endif
*/
/**
* @brief Extracts all spot_ids from metadata into m_spot_id_buffer and purges metadata spotId columns
*
*/
void extract_spotid() {
m_spot_id_buffer.clear();
int sz = m_read_groups.size();
m_spot_id_buffer.resize(sz);
tf::Taskflow taskflow;
taskflow.for_each_index(0, sz, 1, [&](int i) {
m_read_groups[i]->extract_64bit_column(metadata_t::e_spotId, m_spot_id_buffer[i].values, m_spot_id_buffer[i].ext, true);
});
m_executor->run(taskflow).wait();
}
/**
* @brief Purges metadata column
*
* @tparam T
* @param col_index
*/
template<typename T>
void clear_column(unsigned col_index) {
for (auto& rg : m_read_groups) {
rg->template clear_column<T>(col_index);
}
}
/**
* @brief Packs all groups that exceed batch_size limit
* groups with less then 1M spots ignored
*
* @param batch_size
*/
void pack_read_groups(size_t batch_size)
{
size_t total_sz = 0;
unsigned num_candidates = 0;
for (auto& rg : m_read_groups) {
if (rg->m_curr_row > 1e6) {
++num_candidates;
}
total_sz += rg->m_curr_row;
}
if (num_candidates > 0) {
int batch_half = batch_size / 2;
auto limit = ((num_candidates * batch_half) + batch_half) / num_candidates;
for (auto& rg : m_read_groups) {
if (rg->m_curr_row >= limit) {
total_sz -= rg->m_curr_row;
rg->pack_batch();
}
}
}
// Keep packing while total group_size exceeds batch_size * 2
while (total_sz >= batch_size * 2) {
int max_index = 0;
size_t max_value = 0;
for (size_t index = 0; index < m_read_groups.size(); ++index) {
if (m_read_groups[index]->m_curr_row > max_value) {
max_value = m_read_groups[index]->m_curr_row;
max_index = index;
}
}
total_sz -= max_value;
m_read_groups[max_index]->pack_batch();
}
}
} context_t;
#if 0
static char const *Print_ctx_value_t(ctx_value_t const *const self)
{
static char buffer[16384];
rc_t rc = string_printf(buffer, sizeof(buffer), NULL, "pid: { %lu, %lu }, sid: %lu, fid: %u, alc: { %u, %u }, flg: %x", CTX_VALUE_GET_P_ID(*self, 0), CTX_VALUE_GET_P_ID(*self, 1), CTX_VALUE_GET_S_ID(*self), self->fragmentId, self->alignmentCount[0], self->alignmentCount[1], *(self->alignmentCount + sizeof(self->alignmentCount)/sizeof(self->alignmentCount[0])));
if (rc)
return 0;
return buffer;
}
#endif
#if defined(HAS_CTX_VALUE)
static rc_t MMArrayMake(MMArray **rslt, int fd, uint32_t elemSize)
{
MMArray *const self = (MMArray *)calloc(1, sizeof(*self));
if (self == NULL)
return RC(rcExe, rcMemMap, rcConstructing, rcMemory, rcExhausted);
self->elemSize = (elemSize + 3) & ~(3u); /** align to 4 byte **/
self->fd = fd;
*rslt = self;
return 0;
}
#define PERF 0
#define PROT 0
static rc_t MMArrayGet(MMArray *const self, void **const value, uint64_t const element)
{
size_t const chunk = MMA_SUBCHUNK_SIZE * self->elemSize;
unsigned const bin_no = element >> 32;
unsigned const subbin = ((uint32_t)element) >> MMA_NUM_CHUNKS_BITS;
unsigned const in_bin = (uint32_t)element & (MMA_SUBCHUNK_SIZE - 1);
if (bin_no >= sizeof(self->map)/sizeof(self->map[0]))
return RC(rcExe, rcMemMap, rcConstructing, rcId, rcExcessive);
if (self->map[bin_no].submap[subbin].base == NULL) {
off_t const cur_fsize = self->fsize;
off_t const new_fsize = cur_fsize + chunk;
if (ftruncate(self->fd, new_fsize) != 0)
return RC(rcExe, rcFile, rcResizing, rcSize, rcExcessive);
else {
void *const base = mmap(NULL, chunk, PROT_READ|PROT_WRITE,
MAP_FILE|MAP_SHARED, self->fd, cur_fsize);
self->fsize = new_fsize;
if (base == MAP_FAILED) {
PLOGMSG(klogErr, (klogErr, "Failed to construct map for bin $(bin), subbin $(subbin)", "bin=%u,subbin=%u", bin_no, subbin));
return RC(rcExe, rcMemMap, rcConstructing, rcMemory, rcExhausted);
}
else {
#if PERF
static unsigned mapcount = 0;
(void)PLOGMSG(klogInfo, (klogInfo, "Number of mmaps: $(cnt)", "cnt=%u", ++mapcount));
#endif
self->map[bin_no].submap[subbin].base = (uint8_t*)base;
}
}
}
uint8_t *const next = self->map[bin_no].submap[subbin].base;
#if PROT
if (next != self->current) {
void *const current = self->current;
if (current)
mprotect(current, chunk, PROT_NONE);
mprotect(self->current = next, chunk, PROT_READ|PROT_WRITE);
}
#endif
*value = &next[(size_t)in_bin * self->elemSize];
return 0;
}
#if 0
static rc_t MMArrayGetRead(MMArray *const self, void const **const value, uint64_t const element)
{
unsigned const bin_no = element >> 32;
unsigned const subbin = ((uint32_t)element) >> MMA_NUM_CHUNKS_BITS;
unsigned const in_bin = (uint32_t)element & (MMA_SUBCHUNK_SIZE - 1);
if (bin_no >= sizeof(self->map)/sizeof(self->map[0]))
return RC(rcExe, rcMemMap, rcConstructing, rcId, rcExcessive);
if (self->map[bin_no].submap[subbin].base == NULL)
return RC(rcExe, rcMemMap, rcReading, rcId, rcInvalid);
uint8_t *const next = self->map[bin_no].submap[subbin].base;
#if PROT
size_t const chunk = MMA_SUBCHUNK_SIZE * self->elemSize;
if (next != self->current) {
void *const current = self->current;
if (current)
mprotect(current, chunk, PROT_NONE);
mprotect(self->current = next, chunk, PROT_READ);
}
#endif
*value = &next[(size_t)in_bin * self->elemSize];
return 0;
}
#endif
static void MMArrayLock(MMArray *const self)
{
#if PROT
size_t const chunk = MMA_SUBCHUNK_SIZE * self->elemSize;
void *const current = self->current;
self->current = NULL;
if (current)
mprotect(current, chunk, PROT_NONE);
#endif
}
static void MMArrayClear(MMArray *self)
{
size_t const chunk = MMA_SUBCHUNK_SIZE * self->elemSize;
unsigned i;
for (i = 0; i != sizeof(self->map)/sizeof(self->map[0]); ++i) {
unsigned j;
for (j = 0; j != sizeof(self->map[0].submap)/sizeof(self->map[0].submap[0]); ++j) {
if (self->map[i].submap[j].base) {
#if PROT
mprotect(self->map[i].submap[j].base, chunk, PROT_READ|PROT_WRITE);
#endif
memset(self->map[i].submap[j].base, 0, chunk);
#if PROT
mprotect(self->map[i].submap[j].base, chunk, PROT_NONE);
#endif
}
}
}
#if PROT
self->current = NULL;
#endif
}
static void MMArrayWhack(MMArray *self)
{
if ( self == NULL )
{
return;
}
size_t const chunk = MMA_SUBCHUNK_SIZE * self->elemSize;
unsigned i;
for (i = 0; i != sizeof(self->map)/sizeof(self->map[0]); ++i) {
unsigned j;
for (j = 0; j != sizeof(self->map[0].submap)/sizeof(self->map[0].submap[0]); ++j) {
if (self->map[i].submap[j].base)
munmap(self->map[i].submap[j].base, chunk);
}
}
close(self->fd);
free(self);
}
#endif
static rc_t GetKeyIDOld(context_t *const ctx,
queue_rec_t& queue_rec,
char const key[], char const name[], unsigned const namelen)
{
unsigned const keylen = strlen(key);
assert(!ctx->m_read_groups.empty());
auto& rs = *ctx->m_read_groups[ctx->m_emptyGroupIndex];
BAM_Alignment& rec = *queue_rec.alignment;
if (memcmp(key, name, keylen) == 0) {
// qname starts with read group; no append
auto& r = rs.find(name, namelen);
rec.keyId = r.pos;
rec.wasInserted = r.wasInserted;
queue_rec.metadata = r.metadata;
queue_rec.row_id = r.row_id;
} else {
char sbuf[4096];
char *buf = sbuf;
char *hbuf = NULL;
size_t bsize = sizeof(sbuf);
size_t actsize;
if (keylen + namelen + 2 > bsize) {
hbuf = (char*)malloc(bsize = keylen + namelen + 2);
if (hbuf == NULL)
return RC(rcExe, rcName, rcAllocating, rcMemory, rcExhausted);
buf = hbuf;
}
string_printf(buf, bsize, &actsize, "%s\t%.*s", key, (int)namelen, name);
auto& r = rs.find(buf, actsize);
rec.keyId = r.pos;
rec.wasInserted = r.wasInserted;
queue_rec.metadata = r.metadata;
queue_rec.row_id = r.row_id;
if (hbuf)
free(hbuf);
}
return 0;
}
#define USE_ILLUMINA_NAMING_CORRECTION 1
static size_t GetFixedNameLength(char const name[], size_t const namelen)
{
#if USE_ILLUMINA_NAMING_CORRECTION
/*** Check for possible fixes to illumina names ****/
size_t newlen=namelen;
/*** First get rid of possible "/1" "/2" "/3" at the end - violates SAM spec **/
if(newlen > 2 && name[newlen-2] == '/' && (name[newlen-1] == '1' || name[newlen-1] == '2' || name[newlen-1] == '3')){
newlen -=2;
}
if(newlen > 2 && name[newlen-2] == '#' && (name[newlen-1] == '0')){ /*** Now, find "#0" ***/
newlen -=2;
} else if(newlen>10){ /*** find #ACGT ***/
int i=newlen;
for(i--;i>4;i--){ /*** stopping at 4 since the rest of record should still contain :x:y ***/
char a=toupper(name[i]);
if(a != 'A' && a != 'C' && a !='G' && a !='T'){
break;
}
}
if (name[i] == '#'){
switch (newlen-i) { /** allowed values for illumina barcodes :5,6,8 **/
case 5:
case 6:
case 8:
newlen=i;
break;
default:
break;
}
}
}
if(newlen < namelen){ /*** check for :x:y at the end now - to make sure it is illumina **/
int i=newlen;
for(i--;i>0 && isdigit(name[i]);i--){}
if(name[i]==':'){
for(i--;i>0 && isdigit(name[i]);i--){}
if(name[i]==':' && newlen > 0){ /*** some name before :x:y should still exist **/
/*** looks like illumina ***/
return newlen;
}
}
}
#endif
return namelen;
}
static bool platform_cmp(char const platform[], char const test[])
{
unsigned i;
for (i = 0; ; ++i) {
int ch1 = test[i];
int ch2 = toupper(platform[i]);
if (ch1 != ch2)
break;
if (ch1 == 0)
return true;
}
return false;
}
static
INSDC_SRA_platform_id GetINSDCPlatform(BAM_File const *bam, char const name[]) {
if (name) {
BAMReadGroup const *rg;
BAM_FileGetReadGroupByName(bam, name, &rg);
if (rg && rg->platform) {
switch (toupper(rg->platform[0])) {
case 'C':
if (platform_cmp(rg->platform, "COMPLETE GENOMICS"))
return SRA_PLATFORM_COMPLETE_GENOMICS;
if (platform_cmp(rg->platform, "CAPILLARY"))
return SRA_PLATFORM_CAPILLARY;
break;
case 'H':
if (platform_cmp(rg->platform, "HELICOS"))
return SRA_PLATFORM_HELICOS;
break;
case 'I':
if (platform_cmp(rg->platform, "ILLUMINA"))
return SRA_PLATFORM_ILLUMINA;
if (platform_cmp(rg->platform, "IONTORRENT"))
return SRA_PLATFORM_ION_TORRENT;
break;
case 'L':
if (platform_cmp(rg->platform, "LS454"))
return SRA_PLATFORM_454;
break;
case 'N':
if (platform_cmp(name, "NANOPORE"))
return SRA_PLATFORM_OXFORD_NANOPORE;
break;
case 'O':
if (platform_cmp(name, "OXFORD_NANOPORE"))
return SRA_PLATFORM_OXFORD_NANOPORE;
break;
case 'P':
if (platform_cmp(rg->platform, "PACBIO"))
return SRA_PLATFORM_PACBIO_SMRT;
break;
case 'S':
if (platform_cmp(rg->platform, "SOLID"))
return SRA_PLATFORM_ABSOLID;
if (platform_cmp(name, "SANGER"))
return SRA_PLATFORM_CAPILLARY;
break;
default:
break;
}
}
}
return SRA_PLATFORM_UNDEFINED;
}
static
rc_t GetKeyID(context_t *const ctx,
const BAM_File* bam,
queue_rec_t& queue_rec,
char const key[],
char const name[],
size_t const o_namelen)
{
static size_t key_count = 0;
static size_t spot_count = 0;
static size_t last_spot_count = 0;
BAM_Alignment& rec = *queue_rec.alignment;
size_t group_id = ctx->m_read_groups.size();
size_t const namelen = GetFixedNameLength(name, o_namelen);
if (ctx->m_isSingleGroup) {
GetKeyIDOld(ctx, queue_rec, key, name, namelen);
if (++key_count % 10000000 == 0) {
auto& spot_assembly = *ctx->m_read_groups.front();
spdlog::info("Group: '{}', batch memory {:L}, filter memory {:L}", key, spot_assembly.memory_used(), spot_assembly.m_key_filter->memory_used());
}
} else {
auto [it, inserted] = ctx->m_group_map.emplace(key, group_id);
if (!inserted) {
group_id = it.value();
} else {
// Created new read group
if (group_id >= MAX_GROUPS_ALLOWED) {
(void)PLOGMSG(klogErr, (klogErr, "too many read groups: max is $(max)", "max=%d", (int) NUM_ID_SPACES));
return RC(rcExe, rcTree, rcAllocating, rcConstraint, rcViolated);
}
ctx->add_read_group().m_platform = GetINSDCPlatform(bam, key);
}
auto& spot_assembly = *ctx->m_read_groups[group_id];
auto& r = spot_assembly.find(name, namelen);
rec.wasInserted = r.wasInserted;
queue_rec.metadata = r.metadata;
queue_rec.row_id = r.row_id;
rec.platform = spot_assembly.m_platform;
rec.keyId = (((uint64_t)group_id) << GROUPID_SHIFT) | r.pos;
if (++key_count % 10000000 == 0) {
spdlog::info("Group: '{}', batch memory {:L}, filter memory {:L}", key, spot_assembly.memory_used(), spot_assembly.m_key_filter->memory_used());
}
}
spot_count += rec.wasInserted ? 1 : 0;
// Check if read_groups need to be packed
if (spot_count % 10000000 == 0 && spot_count && spot_count != last_spot_count) {
last_spot_count = spot_count;
size_t num_chunks = 0;
// Estimated batch size until 10% of input size is processed
if (ctx->m_calcBatchSize && ctx->m_inputSize) {
BAM_FilePosition end_pos = 0;
BAM_FileGetPosition(bam, &end_pos);
end_pos >>= 16;
size_t chunk_len = end_pos - ctx->m_fileOffset;
ctx->m_processedSize += chunk_len;
ctx->m_fileOffset = end_pos;
if (ctx->m_processedSize) {
num_chunks = (ctx->m_inputSize / ctx->m_processedSize) + 1;
size_t num_spots = num_chunks * spot_count;
ctx->set_key_filter(num_spots);
ctx->m_estimatedBatchSize = min<int>(G.searchBatchSize, num_spots/(ctx->m_executor->num_workers() - 1));
ctx->m_estimatedBatchSize = max<int>(10e6, ctx->m_estimatedBatchSize);
if ((float)ctx->m_processedSize/ctx->m_inputSize > 0.1f) {
ctx->m_calcBatchSize = false;
}
spdlog::info("Current spot_count: {:L}, estimated spot count {:L}, estimated batch size: {:L}", spot_count, num_spots, ctx->m_estimatedBatchSize);
}
}
ctx->pack_read_groups(ctx->m_estimatedBatchSize);
}
return 0;
}
#if defined HAS_CTX_VALUE
static rc_t OpenMMapFile(context_t *const ctx, KDirectory *const dir)
{
int fd;
char fname[4096];
rc_t rc = string_printf(fname, sizeof(fname), NULL, "%s/id2value.%u", G.tmpfs, G.pid);
if (rc)
return rc;
fd = open(fname, O_RDWR|O_TRUNC|O_CREAT, S_IRUSR|S_IWUSR);
if (fd < 0)
return RC(rcExe, rcFile, rcCreating, rcFile, rcNotFound);
unlink(fname);
return MMArrayMake(&ctx->id2value, fd, sizeof(ctx_value_t));
}
#endif
static rc_t TmpfsDirectory(KDirectory **const rslt)
{
KDirectory *dir;
rc_t rc = KDirectoryNativeDir(&dir);
if (rc == 0) {
rc = KDirectoryOpenDirUpdate(dir, rslt, false, "%s", G.tmpfs);
KDirectoryRelease(dir);
}
return rc;
}
static rc_t SetupContext(context_t *ctx, unsigned numfiles)
{
rc_t rc = 0;
// memset(ctx, 0, sizeof(*ctx));
if (G.mode == mode_Archive) {
KDirectory *dir;
size_t fragSize[2];
fragSize[1] = (G.cache_size / 8);
fragSize[0] = fragSize[1] * 4;
rc = TmpfsDirectory(&dir);
#if defined HAS_CTX_VALUE
if (rc == 0)
rc = OpenMMapFile(ctx, dir);
#endif
if (rc == 0)
rc = MemBankMake(&ctx->frags, dir, G.pid, fragSize);
KDirectoryRelease(dir);
}
else if (G.mode == mode_Remap) {
ctx->reset_for_remap();
}
rc = KLoadProgressbar_Make(&ctx->progress[0], 0); if (rc) return rc;
rc = KLoadProgressbar_Make(&ctx->progress[1], 0); if (rc) return rc;
rc = KLoadProgressbar_Make(&ctx->progress[2], 0); if (rc) return rc;
rc = KLoadProgressbar_Make(&ctx->progress[3], 0); if (rc) return rc;
KLoadProgressbar_Append(ctx->progress[0], 100 * numfiles);
ctx->m_estimatedBatchSize = G.searchBatchSize;
ctx->m_key_filter.reset(new fnv_murmur_filter);
ctx->m_executor.reset(new tf::Executor(G.numThreads));
return rc;
}
static void ContextReleaseMemBank(context_t *ctx)
{
MemBankRelease(ctx->frags);
ctx->frags = NULL;
}
static void ContextRelease(context_t *ctx, bool continuing)
{
KLoadProgressbar_Release(ctx->progress[0], true);
KLoadProgressbar_Release(ctx->progress[1], true);
KLoadProgressbar_Release(ctx->progress[2], true);
KLoadProgressbar_Release(ctx->progress[3], true);
#ifdef HAS_CTX_VALUE
if (!continuing)
MMArrayWhack(ctx->id2value);
else
MMArrayClear(ctx->id2value);
#endif
}
static
void COPY_QUAL(uint8_t D[], uint8_t const S[], unsigned const L, bool const R)
{
if (R) {
unsigned i;
unsigned j;
for (i = 0, j = L - 1; i != L; ++i, --j)
D[i] = S[j];
}
else
memmove(D, S, L);
}
static
void COPY_READ(INSDC_dna_text D[], INSDC_dna_text const S[], unsigned const L, bool const R)
{
static INSDC_dna_text const complement[] = {
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , '.', 0 ,
'0', '1', '2', '3', 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 'T', 'V', 'G', 'H', 0 , 0 , 'C',
'D', 0 , 0 , 'M', 0 , 'K', 'N', 0 ,
0 , 0 , 'Y', 'S', 'A', 'A', 'B', 'W',
0 , 'R', 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 'T', 'V', 'G', 'H', 0 , 0 , 'C',
'D', 0 , 0 , 'M', 0 , 'K', 'N', 0 ,
0 , 0 , 'Y', 'S', 'A', 'A', 'B', 'W',
0 , 'R', 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0 ,
0 , 0 , 0 , 0 , 0 , 0 , 0 , 0
};
if (R) {
unsigned i;
unsigned j;
for (i = 0, j = L - 1; i != L; ++i, --j)
D[i] = complement[((uint8_t const *)S)[j]];
}
else
memmove(D, S, L);
}
static KFile *MakeDeferralFile() {
if (G.deferSecondary) {
char tmplate[4096];
int fd;
KFile *f;
KDirectory *d;
size_t nwrit;
KDirectoryNativeDir(&d);
string_printf(tmplate, sizeof(tmplate), &nwrit, "%s/defer.XXXXXX", G.tmpfs);
fd = mkstemp(tmplate);
KDirectoryOpenFileWrite(d, &f, true, tmplate);
KDirectoryRelease(d);
close(fd);
unlink(tmplate);
return f;
}
return NULL;
}
static rc_t OpenBAM(const BAM_File **bam, VDatabase *db, const char bamFile[])
{
rc_t rc = 0;
KFile *defer = MakeDeferralFile();
if (strcmp(bamFile, "/dev/stdin") == 0) {
rc = BAM_FileMake(bam, defer, G.headerText, "/dev/stdin");
}
else {
rc = BAM_FileMake(bam, defer, G.headerText, "%s", bamFile);
}
KFileRelease(defer); /* it was retained by BAM file */
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "Failed to open '$(file)'", "file=%s", bamFile));
}
else if (db) {
KMetadata *dbmeta;
rc = VDatabaseOpenMetadataUpdate(db, &dbmeta);
if (rc == 0) {
KMDataNode *node;
rc = KMetadataOpenNodeUpdate(dbmeta, &node, "BAM_HEADER");
KMetadataRelease(dbmeta);
if (rc == 0) {
char const *header;
size_t size;
rc = BAM_FileGetHeaderText(*bam, &header, &size);
if (rc == 0) {
rc = KMDataNodeWrite(node, header, size);
}
KMDataNodeRelease(node);
}
}
}
return rc;
}
static rc_t VerifyReferences(BAM_File const *bam, Reference const *ref)
{
rc_t rc = 0;
uint32_t n;
unsigned i;
BAM_FileGetRefSeqCount(bam, &n);
for (i = 0; i != n; ++i) {
BAMRefSeq const *refSeq;
BAM_FileGetRefSeq(bam, i, &refSeq);
if (G.refFilter && strcmp(refSeq->name, G.refFilter) != 0)
continue;
rc = ReferenceVerify(ref, refSeq->name, refSeq->length, refSeq->checksum);
if (rc) {
if (GetRCObject(rc) == rcId && GetRCState(rc) == rcUndefined) {
(void)PLOGMSG(klogInfo, (klogInfo, "Reference: '$(name)' is unmapped", "name=%s", refSeq->name));
}
else if (GetRCObject(rc) == rcChecksum && GetRCState(rc) == rcUnequal) {
#if NCBI
(void)PLOGMSG(klogWarn, (klogWarn, "Reference: '$(name)', Length: $(len); checksums do not match", "name=%s,len=%u", refSeq->name, (unsigned)refSeq->length));
#endif
}
else if (GetRCObject(rc) == rcSize && GetRCState(rc) == rcUnequal) {
(void)PLOGMSG(klogWarn, (klogWarn, "Reference: '$(name)', Length: $(len); lengths do not match", "name=%s,len=%u", refSeq->name, (unsigned)refSeq->length));
}
else if (GetRCObject(rc) == rcSize && GetRCState(rc) == rcEmpty) {
(void)PLOGMSG(klogWarn, (klogWarn, "Reference: '$(name)', Length: $(len); fasta file is empty", "name=%s,len=%u", refSeq->name, (unsigned)refSeq->length));
}
else if (GetRCObject(rc) == rcId && GetRCState(rc) == rcNotFound) {
(void)PLOGMSG(klogWarn, (klogWarn, "Reference: '$(name)', Length: $(len); no match found", "name=%s,len=%u", refSeq->name, (unsigned)refSeq->length));
}
else {
(void)PLOGERR(klogWarn, (klogWarn, rc, "Reference: '$(name)', Length: $(len); error", "name=%s,len=%u", refSeq->name, (unsigned)refSeq->length));
}
}
else if (G.onlyVerifyReferences) {
(void)PLOGMSG(klogInfo, (klogInfo, "Reference: '$(name)', Length: $(len); match found", "name=%s,len=%u", refSeq->name, (unsigned)refSeq->length));
}
}
return 0;
}
static uint8_t GetMapQ(BAM_Alignment const *rec)
{
uint8_t mapQ;
BAM_AlignmentGetMapQuality(rec, &mapQ);
return mapQ;
}
#if 0
static bool EditAlignedQualities(uint8_t qual[], bool const hasMismatch[], unsigned readlen)
{
unsigned i;
bool changed = false;
for (i = 0; i < readlen; ++i) {
uint8_t const q_0 = qual[i];
uint8_t const q_1= hasMismatch[i] ? G.alignedQualValue : q_0;
if (q_0 != q_1) {
changed = true;
break;
}
}
if (!changed)
return false;
for (i = 0; i < readlen; ++i) {
uint8_t const q_0 = qual[i];
uint8_t const q_1= hasMismatch[i] ? G.alignedQualValue : q_0;
qual[i] = q_1;
}
return true;
}
#endif
#if 0
static bool EditUnalignedQualities(uint8_t qual[], bool const hasMismatch[], unsigned readlen)
{
unsigned i;
bool changed = false;
for (i = 0; i < readlen; ++i) {
uint8_t const q_0 = qual[i];
uint8_t const q_1 = (q_0 & 0x7F) | (hasMismatch[i] ? 0x80 : 0);
if (q_0 != q_1) {
changed = true;
break;
}
}
if (!changed)
return false;
for (i = 0; i < readlen; ++i) {
uint8_t const q_0 = qual[i];
uint8_t const q_1 = (q_0 & 0x7F) | (hasMismatch[i] ? 0x80 : 0);
qual[i] = q_1;
}
return true;
}
#endif
static
rc_t CheckLimitAndLogError(void)
{
unsigned const count = ++G.errCount;
if (G.maxErrCount > 0 && count > G.maxErrCount) {
(void)PLOGERR(klogErr, (klogErr, SILENT_RC(rcAlign, rcFile, rcReading, rcError, rcExcessive), "Number of errors $(cnt) exceeds limit of $(max): Exiting", "cnt=%u,max=%u", count, G.maxErrCount));
return RC(rcAlign, rcFile, rcReading, rcError, rcExcessive);
}
return 0;
}
static
void RecordNoMatch(char const readName[], char const refName[], uint32_t const refPos)
{
if (G.noMatchLog) {
static uint64_t lpos = 0;
char logbuf[256];
size_t len;
if (string_printf(logbuf, sizeof(logbuf), &len, "%s\t%s\t%u\n", readName, refName, refPos) == 0) {
KFileWrite(G.noMatchLog, lpos, logbuf, len, NULL);
lpos += len;
}
}
}
static LowMatchCounter *lmc = NULL;
static
rc_t LogNoMatch(char const readName[], char const refName[], unsigned rpos, unsigned matches)
{
rc_t const rc = CheckLimitAndLogError();
static unsigned count = 0;
if (lmc == NULL)
lmc = LowMatchCounterMake();
assert(lmc != NULL);
LowMatchCounterAdd(lmc, refName);
++count;
if (rc) {
(void)PLOGMSG(klogInfo, (klogInfo, "This is the last warning; this class of warning occurred $(occurred) times",
"occurred=%u", count));
(void)PLOGMSG(klogErr, (klogErr, "Spot '$(name)' contains too few ($(count)) matching bases to reference '$(ref)' at $(pos)",
"name=%s,ref=%s,pos=%u,count=%u", readName, refName, rpos, matches));
return rc;
}
if (G.maxWarnCount_NoMatch == 0 || count < G.maxWarnCount_NoMatch)
(void)PLOGMSG(klogWarn, (klogWarn, "Spot '$(name)' contains too few ($(count)) matching bases to reference '$(ref)' at $(pos)",
"name=%s,ref=%s,pos=%u,count=%u", readName, refName, rpos, matches));
return 0;
}
struct rlmc_context {
KMDataNode *node;
unsigned node_number;
rc_t rc;
};
static void RecordLowMatchCount(void *Ctx, char const name[], unsigned const count)
{
struct rlmc_context *const ctx = (rlmc_context *)Ctx;
if (ctx->rc == 0) {
KMDataNode *sub = NULL;
ctx->rc = KMDataNodeOpenNodeUpdate(ctx->node, &sub, "LOW_MATCH_COUNT_%u", ++ctx->node_number);
if (ctx->rc == 0) {
uint32_t const count_temp = count;
ctx->rc = KMDataNodeWriteAttr(sub, "REFNAME", name);
if (ctx->rc == 0)
ctx->rc = KMDataNodeWriteB32(sub, &count_temp);
KMDataNodeRelease(sub);
}
}
}
static rc_t RecordLowMatchCounts(KMDataNode *const node)
{
struct rlmc_context ctx;
assert(lmc != NULL);
if (node) {
ctx.node = node;
ctx.node_number = 0;
ctx.rc = 0;
LowMatchCounterEach(lmc, &ctx, RecordLowMatchCount);
}
return ctx.rc;
}
#if 0
static
rc_t LogDupConflict(char const readName[])
{
rc_t const rc = CheckLimitAndLogError();
static unsigned count = 0;
++count;
if (rc) {
(void)PLOGMSG(klogInfo, (klogInfo, "This is the last warning; this class of warning occurred $(occurred) times",
"occurred=%u", count));
(void)PLOGERR(klogWarn, (klogWarn, SILENT_RC(rcApp, rcFile, rcReading, rcData, rcInconsistent),
"Spot '$(name)' is both a duplicate and NOT a duplicate!",
"name=%s", readName));
}
else if (G.maxWarnCount_DupConflict == 0 || count < G.maxWarnCount_DupConflict)
(void)PLOGERR(klogWarn, (klogWarn, SILENT_RC(rcApp, rcFile, rcReading, rcData, rcInconsistent),
"Spot '$(name)' is both a duplicate and NOT a duplicate!",
"name=%s", readName));
return rc;
}
#endif
static char const *const CHANGED[] = {
"FLAG changed",
"QUAL changed",
"SEQ changed",
"record made unaligned",
"record made unfragmented",
"mate alignment lost",
"record discarded",
"reference name changed",
"CIGAR changed"
};
#define FLAG_CHANGED (0)
#define QUAL_CHANGED (1)
#define SEQ_CHANGED (2)
#define MAKE_UNALIGNED (3)
#define MAKE_UNFRAGMENTED (4)
#define MATE_LOST (5)
#define DISCARDED (6)
#define REF_NAME_CHANGED (7)
#define CIGAR_CHANGED (8)
static char const *const REASONS[] = {
/* FLAG changed */
"0x400 and 0x200 both set", /* 0 */
"conflicting PCR Dup flags", /* 1 */
"primary alignment already exists", /* 2 */
"was already recorded as unaligned", /* 3 */
/* QUAL changed */
"original quality used", /* 4 */
"unaligned colorspace", /* 5 */
"aligned bases", /* 6 */
"unaligned bases", /* 7 */
"reversed", /* 8 */
/* unaligned */
"low MAPQ", /* 9 */
"low match count", /* 10 */
"missing alignment info", /* 11 */
"missing reference position", /* 12 */
"invalid alignment info", /* 13 */
"invalid reference position", /* 14 */
"invalid reference", /* 15 */
"unaligned reference", /* 16 */
"unknown reference", /* 17 */
"hard-clipped colorspace", /* 18 */
/* unfragmented */
"missing fragment info", /* 19 */
"too many fragments", /* 20 */
/* mate info lost */
"invalid mate reference", /* 21 */
"missing mate alignment info", /* 22 */
"unknown mate reference", /* 23 */
/* discarded */
"conflicting PCR duplicate", /* 24 */
"conflicting fragment info", /* 25 */
"reference is skipped", /* 26 */
/* reference name changed */
"reference was named more than once", /* 27 */
/* CIGAR changed */
"alignment overhanging end of reference", /* 28 */
/* discarded */
"hard-clipped secondary alignment", /* 29 */
"low-matching secondary alignment", /* 30 */
};
static struct {
unsigned what, why;
} const CHANGES[] = {
{FLAG_CHANGED, 0},
{FLAG_CHANGED, 1},
{FLAG_CHANGED, 2},
{FLAG_CHANGED, 3},
{QUAL_CHANGED, 4},
{QUAL_CHANGED, 5},
{QUAL_CHANGED, 6},
{QUAL_CHANGED, 7},
{QUAL_CHANGED, 8},
{SEQ_CHANGED, 8},
{MAKE_UNALIGNED, 9},
{MAKE_UNALIGNED, 10},
{MAKE_UNALIGNED, 11},
{MAKE_UNALIGNED, 12},
{MAKE_UNALIGNED, 13},
{MAKE_UNALIGNED, 14},
{MAKE_UNALIGNED, 15},
{MAKE_UNALIGNED, 16},
{MAKE_UNALIGNED, 17},
{MAKE_UNALIGNED, 18},
{MAKE_UNFRAGMENTED, 19},
{MAKE_UNFRAGMENTED, 20},
{MATE_LOST, 21},
{MATE_LOST, 22},
{MATE_LOST, 23},
{DISCARDED, 24},
{DISCARDED, 25},
{DISCARDED, 26},
{DISCARDED, 17},
{REF_NAME_CHANGED, 27},
{CIGAR_CHANGED, 28},
{DISCARDED, 29},
{DISCARDED, 30},
};
#define NUMBER_OF_CHANGES ((unsigned)(sizeof(CHANGES)/sizeof(CHANGES[0])))
static unsigned change_counter[NUMBER_OF_CHANGES];
static void LOG_CHANGE(unsigned const change)
{
++change_counter[change];
}
static void PrintChangeReport(void)
{
unsigned i;
for (i = 0; i != NUMBER_OF_CHANGES; ++i) {
if (change_counter[i] > 0) {
char const *const what = CHANGED[CHANGES[i].what];
char const *const why = REASONS[CHANGES[i].why];
PLOGMSG(klogInfo, (klogInfo, "$(what) $(times) times because $(reason)", "what=%s,reason=%s,times=%u", what, why, change_counter[i]));
}
}
}
static rc_t RecordChange(KMDataNode *const node,
char const node_name[],
unsigned const node_number,
char const what[],
char const why[],
unsigned const count)
{
KMDataNode *sub = NULL;
rc_t const rc_sub = KMDataNodeOpenNodeUpdate(node, &sub, "%s_%u", node_name, node_number);
if (rc_sub) return rc_sub;
{
uint32_t const count_temp = count;
rc_t const rc_attr1 = KMDataNodeWriteAttr(sub, "change", what);
rc_t const rc_attr2 = KMDataNodeWriteAttr(sub, "reason", why);
rc_t const rc_value = KMDataNodeWriteB32(sub, &count_temp);
KMDataNodeRelease(sub);
if (rc_attr1) return rc_attr1;
if (rc_attr2) return rc_attr2;
if (rc_value) return rc_value;
return 0;
}
}
static rc_t RecordChanges(KMDataNode *const node, char const name[])
{
if (node) {
unsigned i;
unsigned j = 0;
for (i = 0; i != NUMBER_OF_CHANGES; ++i) {
if (change_counter[i] > 0) {
char const *const what = CHANGED[CHANGES[i].what];
char const *const why = REASONS[CHANGES[i].why];
rc_t const rc = RecordChange(node, name, ++j, what, why, change_counter[i]);
if (rc) return rc;
}
}
}
return 0;
}
#define FLAG_CHANGED_400_AND_200 do { LOG_CHANGE( 0); } while(0)
#define FLAG_CHANGED_PCR_DUP do { LOG_CHANGE( 1); } while(0)
#define FLAG_CHANGED_PRIMARY_DUP do { LOG_CHANGE( 2); } while(0)
#define FLAG_CHANGED_WAS_UNALIGNED do { LOG_CHANGE( 3); } while(0)
#define QUAL_CHANGED_OQ do { LOG_CHANGE( 4); } while(0)
#define QUAL_CHANGED_UNALIGNED_CS do { LOG_CHANGE( 5); } while(0)
#define QUAL_CHANGED_ALIGNED_EDIT do { LOG_CHANGE( 6); } while(0)
#define QUAL_CHANGED_UNALIGN_EDIT do { LOG_CHANGE( 7); } while(0)
#define QUAL_CHANGED_REVERSED do { LOG_CHANGE( 8); } while(0)
#define SEQ__CHANGED_REV_COMP do { LOG_CHANGE( 9); } while(0)
#define UNALIGNED_LOW_MAPQ do { LOG_CHANGE(10); } while(0)
#define UNALIGNED_LOW_MATCH_COUNT do { LOG_CHANGE(11); } while(0)
#define UNALIGNED_MISSING_INFO do { LOG_CHANGE(12); } while(0)
#define UNALIGNED_MISSING_REF_POS do { LOG_CHANGE(13); } while(0)
#define UNALIGNED_INVALID_INFO do { LOG_CHANGE(14); } while(0)
#define UNALIGNED_INVALID_REF_POS do { LOG_CHANGE(15); } while(0)
#define UNALIGNED_INVALID_REF do { LOG_CHANGE(16); } while(0)
#define UNALIGNED_UNALIGNED_REF do { LOG_CHANGE(17); } while(0)
#define UNALIGNED_UNKNOWN_REF do { LOG_CHANGE(18); } while(0)
#define UNALIGNED_HARD_CLIPPED_CS do { LOG_CHANGE(19); } while(0)
#define UNFRAGMENT_MISSING_INFO do { LOG_CHANGE(20); } while(0)
#define UNFRAGMENT_TOO_MANY do { LOG_CHANGE(21); } while(0)
#define MATE_INFO_LOST_INVALID do { LOG_CHANGE(22); } while(0)
#define MATE_INFO_LOST_MISSING do { LOG_CHANGE(23); } while(0)
#define MATE_INFO_LOST_UNKNOWN_REF do { LOG_CHANGE(24); } while(0)
#define DISCARD_PCR_DUP do { LOG_CHANGE(25); } while(0)
#define DISCARD_BAD_FRAGMENT_INFO do { LOG_CHANGE(26); } while(0)
#define DISCARD_SKIP_REFERENCE do { LOG_CHANGE(27); } while(0)
#define DISCARD_UNKNOWN_REFERENCE do { LOG_CHANGE(28); } while(0)
#define RENAMED_REFERENCE do { LOG_CHANGE(29); } while(0)
#define OVERHANGING_ALIGNMENT do { LOG_CHANGE(30); } while(0)
#define DISCARD_HARDCLIP_SECONDARY do { LOG_CHANGE(31); } while(0)
#define DISCARD_BAD_SECONDARY do { LOG_CHANGE(32); } while(0)
static bool isHardClipped(unsigned const ops, uint32_t const cigar[/* ops */])
{
unsigned i;
for (i = 0; i < ops; ++i) {
uint32_t const op = cigar[i];
int const code = op & 0x0F;
if (code == 5)
return true;
}
return false;
}
static rc_t FixOverhangingAlignment(KDataBuffer *cigBuf, uint32_t *opCount, uint32_t refPos, uint32_t refLen, uint32_t readlen)
{
uint32_t const *cigar = (uint32_t*)cigBuf->base;
int refend = refPos;
int seqpos = 0;
unsigned i;
for (i = 0; i < *opCount; ++i) {
uint32_t const op = cigar[i];
int const len = op >> 4;
int const code = op & 0x0F;
switch (code) {
case 0: /* M */
case 7: /* = */
case 8: /* X */
seqpos += len;
refend += len;
break;
case 2: /* D */
case 3: /* N */
refend += len;
break;
case 1: /* I */
case 4: /* S */
case 9: /* B */
seqpos += len;
default:
break;
}
if (refend > refLen) {
int const chop = refend - refLen;
int const newlen = len - chop;
int const left = seqpos - chop;
if (left * 2 > readlen) {
int const clip = readlen - left;
rc_t rc;
*opCount = i + 2;
rc = KDataBufferResize(cigBuf, *opCount);
if (rc) return rc;
((uint32_t *)cigBuf->base)[i ] = (newlen << 4) | code;
((uint32_t *)cigBuf->base)[i+1] = (clip << 4) | 4;
OVERHANGING_ALIGNMENT;
break;
}
}
}
return 0;
}
static context_t GlobalContext;
#ifdef NEW_QUEUE
static ReaderWriterQueue<queue_rec_t> rw_queue{1024};
atomic<bool> rw_done{false};
#else
static KQueue *bamq;
#endif
static KThread *bamread_thread;
static rc_t BAM_FileReadDetached(BAM_File const *self, BAM_Alignment **rec)
{
BAM_Alignment const *crec = NULL;
rc_t const rc = BAM_FileRead2(self, &crec);
if (rc == 0) {
if ((*rec = BAM_AlignmentDetach(crec)) != NULL)
return 0;
return RC(rcAlign, rcFile, rcReading, rcMemory, rcExhausted);
}
BAM_AlignmentRelease(crec);
return rc;
}
static rc_t run_bamread_thread(const KThread *self, void *const file)
{
rc_t rc = 0;
size_t NR = 0;
auto bam = (const BAM_File*)file;
while (rc == 0) {
if (rw_done)
break;
BAM_Alignment *rec = NULL;
++NR;
rc = BAM_FileReadDetached(bam, &rec);
if ((int)GetRCObject(rc) == rcRow && (int)GetRCState(rc) == rcEmpty) {
rc = CheckLimitAndLogError();
continue;
}
if ((int)GetRCObject(rc) == rcRow && (int)GetRCState(rc) == rcNotFound) {
/* EOF */
rc = 0;
--NR;
break;
}
if (rc) break;
#if defined(NEW_QUEUE)
queue_rec_t queue_rec;
#else
queue_rec_t* queue_rec = new queue_rec_t;
#endif
{
static char const dummy[] = "";
char const *spotGroup;
char const *name;
size_t namelen;
BAM_AlignmentGetReadName2(rec, &name, &namelen);
BAM_AlignmentGetReadGroupName(rec, &spotGroup);
#if defined(NEW_QUEUE)
queue_rec.alignment = rec;
queue_rec.metadata = nullptr;
rc = GetKeyID(&GlobalContext, bam, queue_rec, spotGroup ? spotGroup : dummy, name, namelen);
#else
queue_rec->alignment = rec;
queue_rec->metadata = nullptr;
rc = GetKeyID(&GlobalContext, bam, *queue_rec, spotGroup ? spotGroup : dummy, name, namelen);
#endif
if (rc) break;
}
for ( ; ; ) {
#ifdef NEW_QUEUE
if (rw_queue.try_enqueue(move(queue_rec))) {
break;
}
if (rw_done)
break;
#else
timeout_t tm;
TimeoutInit(&tm, 1000);
rc = KQueuePush(bamq, queue_rec, &tm);
if (rc == 0 || (int)GetRCObject(rc) != rcTimeout)
break;
#endif
}
}
#ifndef NEW_QUEUE
KQueueSeal(bamq);
#else
rw_done.store(true);
#endif
if (rc) {
(void)LOGERR(klogErr, rc, "bamread_thread done");
}
else {
(void)PLOGMSG(klogInfo, (klogInfo, "bamread_thread done; read $(NR) records", "NR=%lu", NR));
}
return rc;
}
/* call on main thread only */
#ifdef NEW_QUEUE
static queue_rec_t const getNextRecord(BAM_File const *const bam, rc_t *const rc)
#else
static queue_rec_t* const getNextRecord(BAM_File const *const bam, rc_t *const rc)
#endif
{
#ifdef NEW_QUEUE
queue_rec_t queue_rec = {nullptr, nullptr};
#else
queue_rec_t* queue_rec = nullptr;
#endif
#ifndef NEW_QUEUE
if (bamq == NULL) {
*rc = KQueueMake(&bamq, 4096);
if (*rc) return queue_rec;
*rc = KThreadMake(&bamread_thread, run_bamread_thread, (void *)bam);
if (*rc) {
KQueueRelease(bamq);
bamq = NULL;
return queue_rec;
}
}
#endif
static size_t dequeued = 0;
while (*rc == 0 && (*rc = Quitting()) == 0) {
//BAM_Alignment const *rec = NULL;
#ifdef NEW_QUEUE
if (rw_queue.try_dequeue(queue_rec)) {
++dequeued;
return queue_rec;
}
if (rw_done.load())
break;
#else
timeout_t tm;
TimeoutInit(&tm, 10000);
*rc = KQueuePop(bamq, (void **)&queue_rec, &tm);
if (*rc == 0) {
return queue_rec; // this is the normal return
}
if ((int)GetRCObject(*rc) == rcTimeout)
*rc = 0;
else {
if ((int)GetRCObject(*rc) == rcData && (int)GetRCState(*rc) == rcDone)
(void)LOGMSG(klogDebug, "KQueuePop Done");
else
(void)PLOGERR(klogWarn, (klogWarn, *rc, "KQueuePop Error", NULL));
}
#endif
}
spdlog::info("Dequeued: {:L}", dequeued);
rw_done = true;
{
rc_t rc2 = 0;
KThreadWait(bamread_thread, &rc2);
if (rc2 != 0)
*rc = rc2; // return the rc from the reader thread
}
KThreadRelease(bamread_thread);
bamread_thread = NULL;
#ifndef NEW_QUEUE
KQueueRelease(bamq);
bamq = NULL;
#endif
return queue_rec;
}
static void getSpotGroup(BAM_Alignment const *const rec, char spotGroup[])
{
char const *rgname;
BAM_AlignmentGetReadGroupName(rec, &rgname);
if (rgname)
strcpy(spotGroup, rgname);
else
spotGroup[0] = '\0';
}
static char const *getLinkageGroup(BAM_Alignment const *const rec)
{
static char linkageGroup[1024];
char const *BX = NULL;
char const *CB = NULL;
char const *UB = NULL;
linkageGroup[0] = '\0';
BAM_AlignmentGetLinkageGroup(rec, &BX, &CB, &UB);
if (BX == NULL) {
if (CB != NULL && UB != NULL) {
unsigned const cblen = strlen(CB);
unsigned const ublen = strlen(UB);
if (cblen + ublen + 8 < sizeof(linkageGroup)) {
memmove(&linkageGroup[ 0], "CB:", 3);
memmove(&linkageGroup[ 3], CB, cblen);
memmove(&linkageGroup[cblen + 3], "|UB:", 4);
memmove(&linkageGroup[cblen + 7], UB, ublen + 1);
}
}
}
else {
unsigned const bxlen = strlen(BX);
if (bxlen + 1 < sizeof(linkageGroup))
memmove(linkageGroup, BX, bxlen + 1);
}
return linkageGroup;
}
static rc_t ProcessBAM(char const bamFile[], context_t *ctx, VDatabase *db,
/* data outputs */
Reference *ref, Sequence *seq, Alignment *align,
/* output parameters */
bool *had_alignments, bool *had_sequences)
{
const BAM_File *bam;
const BAM_Alignment *rec;
#if defined(NEW_QUEUE)
queue_rec_t queue_rec;
#else
queue_rec_t* queue_rec;
#endif
KDataBuffer buf;
KDataBuffer fragBuf;
KDataBuffer cigBuf;
rc_t rc;
const BAMRefSeq *refSeq = NULL;
int32_t lastRefSeqId = -1;
bool wasRenamed = false;
size_t rsize;
uint64_t keyId = 0;
uint64_t reccount = 0;
char spotGroup[512];
size_t namelen;
float progress = 0.0;
unsigned warned = 0;
long fcountBoth=0;
long fcountOne=0;
int skipRefSeqID = -1;
int unmapRefSeqId = -1;
uint64_t recordsRead = 0;
uint64_t recordsProcessed = 0;
uint64_t filterFlagConflictRecords=0; /*** counts number of conflicts between flags 0x400 and 0x200 ***/
#define MAX_WARNINGS_FLAG_CONFLICT 10000 /*** maximum errors to report ***/
bool isColorSpace = false;
bool isNotColorSpace = G.noColorSpace;
char alignGroup[32];
size_t alignGroupLen;
AlignmentRecord data;
KDataBuffer seqBuffer;
KDataBuffer qualBuffer;
SequenceRecord srec;
SequenceRecordStorage srecStorage;
/* setting up buffers */
memset(&data, 0, sizeof(data));
memset(&srec, 0, sizeof(srec));
srec.ti = srecStorage.ti;
srec.readStart = srecStorage.readStart;
srec.readLen = srecStorage.readLen;
srec.orientation = srecStorage.orientation;
srec.is_bad = srecStorage.is_bad;
srec.alignmentCount = srecStorage.alignmentCount;
srec.aligned = srecStorage.aligned;
srec.cskey = srecStorage. cskey;
rc = OpenBAM(&bam, db, bamFile);
if (rc) return rc;
if (!G.noVerifyReferences && ref != NULL) {
rc = VerifyReferences(bam, ref);
if (G.onlyVerifyReferences) {
BAM_FileRelease(bam);
return rc;
}
}
BAM_FileGetPosition(bam, &ctx->m_fileOffset);
ctx->m_fileOffset >>= 16;
{
uint32_t rgcount;
BAM_FileGetReadGroupCount(bam, &rgcount);
ctx->m_isSingleGroup = rgcount >= MAX_GROUPS_ALLOWED; // TODO
if (ctx->m_isSingleGroup && ctx->m_emptyGroupIndex == -1) {
ctx->add_read_group();
ctx->m_emptyGroupIndex = ctx->m_read_groups.size() - 1;
}
for (unsigned rgi = 0; rgi != rgcount; ++rgi) {
BAMReadGroup const *rg;
BAM_FileGetReadGroup(bam, rgi, &rg);
if (rg && rg->platform && platform_cmp(rg->platform, "CAPILLARY")) {
G.hasTI = true;
break;
}
}
}
/* setting up more buffers */
rc = KDataBufferMake(&cigBuf, 32, 0);
if (rc)
return rc;
rc = KDataBufferMake(&fragBuf, 8, 1024);
if (rc)
return rc;
rc = KDataBufferMake(&buf, 16, 0);
if (rc)
return rc;
rc = KDataBufferMake(&seqBuffer, 8, 4096);
if (rc)
return rc;
rc = KDataBufferMake(&qualBuffer, 8, 4096);
if (rc)
return rc;
if (rc == 0) {
(void)PLOGMSG(klogInfo, (klogInfo, "Loading '$(file)'", "file=%s", bamFile));
}
spdlog::stopwatch sw;
uint64_t primaryId[2];
std::optional<uint8_t> opt_frag_len[2];
std::optional<bool> opt_pcr_dup;
std::optional<bool> opt_is_primary;
std::optional<bool> opt_is_unmated;
#ifdef NEW_QUEUE
//while (rw_queue.pop()); // clear queue
rw_done = false;
auto _rc = KThreadMake(&bamread_thread, run_bamread_thread, (void *)bam);
if (_rc) {
return 0;
}
#endif
size_t new_spots = 0;
string prev_rec;
#if defined(NEW_QUEUE)
while (true) {
queue_rec = getNextRecord(bam, &rc);
rec = queue_rec.alignment;
#else
while ((queue_rec = getNextRecord(bam, &rc)) != NULL) {
rec = queue_rec->alignment;
#endif
if (rec == nullptr)
break;
bool aligned;
uint32_t readlen;
uint16_t flags;
int64_t rpos=0;
char *seqDNA;
#ifdef HAS_CTX_VALUE
ctx_value_t *value;
#endif
bool wasInserted;
int32_t refSeqId=-1;
uint8_t *qual;
bool mated;
const char *name;
char cskey = 0;
bool originally_aligned;
bool isPrimary;
uint32_t opCount;
bool hasCG = false;
uint64_t ti = 0;
uint32_t csSeqLen = 0;
int lpad = 0;
int rpad = 0;
bool hardclipped = false;
bool revcmp = false;
unsigned readNo = 0;
bool wasPromoted = false;
char const *barCode = NULL;
char const *linkageGroup;
keyId = rec->keyId;
wasInserted = rec->wasInserted;
if (wasInserted)
++new_spots;
#ifndef NO_METADATA
//uint64_t row_id = keyId & 0xffffffffff; //(64 - MAX_GROUP_BITS) bits
#if defined(NEW_QUEUE)
auto& metadata = *queue_rec.metadata;
const uint64_t row_id = queue_rec.row_id;
if (metadata.need_optimize) {
spdlog::stopwatch sw;
metadata.memory_used = metadata.Optimize();
spdlog::info("Metadata memory {:L}", metadata.memory_used);
spdlog::info("Metadata Optimize {:.3} sec", sw);
metadata.need_optimize = false;
}
#else
auto& metadata = *queue_rec->metadata;
const uint64_t row_id = queue_rec->row_id;
#endif
primaryId[0] = 0;
primaryId[1] = 0;
opt_pcr_dup.reset();
opt_is_primary.reset();
opt_is_unmated.reset();
opt_frag_len[0].reset();
opt_frag_len[1].reset();
#endif
++recordsRead;
if (recordsRead % 10000000 == 0) {
{
float const new_value = BAM_FileGetProportionalPosition(bam) * 100.0;
float const delta = new_value - progress;
if (delta > 1.0) {
KLoadProgressbar_Process(ctx->progress[0], delta, false);
progress = new_value;
}
}
spdlog::info("Keys {:L}, time: {:.3} sec, memory: {:L}", recordsRead, sw, getCurrentRSS());
sw.reset();
/*
for (auto& gr : ctx->m_read_groups) {
int idx = 0;
for (auto& b : gr->m_batches) {
if (b->m_data_ready && b->m_data_saved == false) {
string fname = fmt::format("{}.{}.batch", gr->m_group_id, idx);
bm::file_save_svector(*b->m_data, fname);
b->m_data_saved = true;
}
++idx;
}
}
*/
}
BAM_AlignmentGetReadName2(rec, &name, &namelen);
#ifdef HAS_CTX_VALUE
rc = MMArrayGet(ctx->id2value, (void **)&value, keyId);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "MMArrayGet: failed on id '$(id)'", "id=%u", keyId));
goto LOOP_END;
}
#endif
linkageGroup = getLinkageGroup(rec);
if (!G.noColorSpace) {
if (BAM_AlignmentHasColorSpace(rec)) {
if (isNotColorSpace) {
MIXED_BASE_AND_COLOR:
rc = RC(rcApp, rcFile, rcReading, rcData, rcInconsistent);
(void)PLOGERR(klogErr, (klogErr, rc, "File '$(file)' contains base space and color space", "file=%s", bamFile));
goto LOOP_END;
}
/* COLORSPACE is disabled!
* ctx->isColorSpace = isColorSpace = true; */
}
else if (isColorSpace)
goto MIXED_BASE_AND_COLOR;
else
isNotColorSpace = true;
}
BAM_AlignmentGetFlags(rec, &flags);
originally_aligned = (flags & BAMFlags_SelfIsUnmapped) == 0;
aligned = originally_aligned;
mated = false;
if (flags & BAMFlags_WasPaired) {
if ((flags & BAMFlags_IsFirst) != 0)
readNo |= 1;
if ((flags & BAMFlags_IsSecond) != 0)
readNo |= 2;
switch (readNo) {
case 1:
case 2:
mated = true;
break;
case 0:
if ((warned & 1) == 0) {
(void)LOGMSG(klogWarn, "Spots without fragment info have been encountered");
warned |= 1;
}
UNFRAGMENT_MISSING_INFO;
break;
case 3:
if ((warned & 2) == 0) {
(void)LOGMSG(klogWarn, "Spots with more than two fragments have been encountered");
warned |= 2;
}
UNFRAGMENT_TOO_MANY;
break;
}
}
if (!mated)
readNo = 1;
isPrimary = (flags & (BAMFlags_IsNotPrimary|BAMFlags_IsSupplemental)) == 0 ? true : false;
#ifndef NO_METADATA
if (G.deferSecondary && !isPrimary && aligned) {
if (wasInserted) {
isPrimary = true;
wasPromoted = true;
} else {
primaryId[readNo - 1] = metadata.get<u64_t>(metadata_t::E_PRIM_ID[readNo - 1]).get_no_check(row_id);
if (primaryId[readNo - 1] == 0) {
/* promote to primary alignment */
isPrimary = true;
wasPromoted = true;
}
}
}
#else
if (G.deferSecondary && !isPrimary && aligned && CTX_VALUE_GET_P_ID(*value, readNo - 1) == 0) {
/* promote to primary alignment */
isPrimary = true;
wasPromoted = true;
}
#endif
if (ctx->m_isSingleGroup)
getSpotGroup(rec, spotGroup);
if (wasInserted) {
if (G.mode == mode_Remap) {
(void)PLOGERR(klogErr, (klogErr, rc = RC(rcApp, rcFile, rcReading, rcData, rcInconsistent),
"Spot '$(name)' is a new spot, not a remapping",
"name=%s", name));
goto LOOP_END;
}
/* first time spot is seen */
/* need to make sure that every goto LOOP_END */
/* above this point is with rc != 0 */
/* else this structure won't get initialized */
#ifndef NO_METADATA
opt_is_unmated = !mated;
if (!mated)
metadata.get<bit_t>(metadata_t::e_unmated).set(row_id);
if (isPrimary || G.assembleWithSecondary || G.deferSecondary) {
opt_pcr_dup = flags & BAMFlags_IsDuplicate;
if (flags & BAMFlags_IsDuplicate)
metadata.get<bit_t>(metadata_t::e_pcr_dup).set(row_id);
if (ctx->m_isSingleGroup)
metadata.get<u16_t>(metadata_t::e_platform).set(row_id, GetINSDCPlatform(bam, spotGroup));
metadata.get<bit_t>(metadata_t::e_primary_is_set).set(row_id);
opt_is_primary = true;
}
#endif
#ifdef HAS_CTX_VALUE
memset(value, 0, sizeof(*value));
value->unmated = !mated;
if (isPrimary || G.assembleWithSecondary || G.deferSecondary) {
value->pcr_dup = (flags & BAMFlags_IsDuplicate) == 0 ? 0 : 1;
value->platform = rec->platform;//GetINSDCPlatform(bam, spotGroup);
//assert(value->platform == rec.platform);
value->primary_is_set = 1;
}
#endif
}
if (!isPrimary && G.noSecondary)
goto LOOP_END;
rc = BAM_AlignmentCGReadLength(rec, &readlen);
if (rc != 0 && GetRCState(rc) != rcNotFound) {
(void)LOGERR(klogErr, rc, "Invalid CG data");
goto LOOP_END;
}
if (rc == 0) {
hasCG = true;
BAM_AlignmentGetCigarCount(rec, &opCount);
rc = KDataBufferResize(&cigBuf, opCount * 2 + 5);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize CIGAR buffer");
goto LOOP_END;
}
rc = AlignmentRecordInit(&data, readlen);
if (rc == 0)
rc = KDataBufferResize(&buf, readlen);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize record buffer");
goto LOOP_END;
}
seqDNA = (char*)buf.base;
qual = (uint8_t *)&seqDNA[readlen];
rc = BAM_AlignmentGetCGSeqQual(rec, seqDNA, qual);
if (rc == 0) {
rc = BAM_AlignmentGetCGCigar(rec, (uint32_t*)cigBuf.base, cigBuf.elem_count, &opCount);
}
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to read CG data");
goto LOOP_END;
}
data.data.align_group.elements = 0;
data.data.align_group.buffer = alignGroup;
if (BAM_AlignmentGetCGAlignGroup(rec, alignGroup, sizeof(alignGroup), &alignGroupLen) == 0)
data.data.align_group.elements = alignGroupLen;
}
else {
/* normal flow i.e. NOT CG */
uint32_t const *tmp;
/* resize buffers */
BAM_AlignmentGetReadLength(rec, &readlen);
BAM_AlignmentGetRawCigar(rec, &tmp, &opCount);
rc = KDataBufferResize(&cigBuf, opCount);
assert(rc == 0);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize CIGAR buffer");
goto LOOP_END;
}
memmove(cigBuf.base, tmp, opCount * sizeof(uint32_t));
hardclipped = isHardClipped(opCount, (const uint32_t*)cigBuf.base);
if (hardclipped) {
if (isPrimary && !wasPromoted) {
/* when we promote a secondary to primary and it is hardclipped, we want to "fix" it */
if (!G.acceptHardClip) {
rc = RC(rcApp, rcFile, rcReading, rcConstraint, rcViolated);
(void)PLOGERR(klogErr, (klogErr, rc, "File '$(file)' contains hard clipped primary alignments", "file=%s", bamFile));
goto LOOP_END;
}
}
else if (!G.acceptHardClip) { /* convert to soft clip */
uint32_t *const cigar = (uint32_t*)cigBuf.base;
uint32_t const lOp = cigar[0];
uint32_t const rOp = cigar[opCount - 1];
lpad = (lOp & 0xF) == 5 ? (lOp >> 4) : 0;
rpad = (rOp & 0xF) == 5 ? (rOp >> 4) : 0;
if (lpad + rpad == 0) {
rc = RC(rcApp, rcFile, rcReading, rcData, rcInvalid);
(void)PLOGERR(klogErr, (klogErr, rc, "File '$(file)' contains invalid CIGAR", "file=%s", bamFile));
goto LOOP_END;
}
if (lpad != 0) {
uint32_t const new_lOp = (((uint32_t)lpad) << 4) | 4;
cigar[0] = new_lOp;
}
if (rpad != 0) {
uint32_t const new_rOp = (((uint32_t)rpad) << 4) | 4;
cigar[opCount - 1] = new_rOp;
}
}
}
if (G.deferSecondary && !isPrimary) {
/*** try to see if hard-clipped secondary alignment can be salvaged **/
auto l = readlen + lpad + rpad;
if (l < 256) {
#ifndef NO_METADATA
uint8_t frag_len = 0;
if (wasInserted == false) {
frag_len = metadata.get<u16_t>(metadata_t::E_FRAG_LEN[readNo - 1]).get_no_check(row_id);
opt_frag_len[readNo - 1] = frag_len;
}
#if defined HAS_CTX_VALUE
if (frag_len != value->fragment_len[readNo - 1]) {
spdlog::error("Inconsistent fragment_len");
throw runtime_error("Inconsistent fragment_len");
}
#endif
#else
auto frag_len = value->fragment_len[readNo -1];
#endif
if ( l < frag_len) {
rc = KDataBufferResize(&cigBuf, opCount + 1);
assert(rc == 0);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize CIGAR buffer");
goto LOOP_END;
}
if (rpad > 0 && lpad == 0) {
uint32_t *const cigar = (uint32_t*)cigBuf.base;
lpad = frag_len - readlen - rpad;
memmove(cigar + 1, cigar, opCount * sizeof(*cigar));
cigar[0] = (uint32_t)((lpad << 4) | 4);
}
else {
uint32_t *const cigar = (uint32_t*)cigBuf.base;
rpad += frag_len - readlen - lpad;
cigar[opCount] = (uint32_t)((rpad << 4) | 4);
}
opCount++;
}
}
}
rc = AlignmentRecordInit(&data, readlen + lpad + rpad);
assert(rc == 0);
if (rc == 0)
rc = KDataBufferResize(&buf, readlen + lpad + rpad);
assert(rc == 0);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize record buffer");
goto LOOP_END;
}
seqDNA = (char*)buf.base;
qual = (uint8_t *)&seqDNA[(readlen | csSeqLen) + lpad + rpad];
memset(seqDNA, 'N', (readlen | csSeqLen) + lpad + rpad);
memset(qual, 0, (readlen | csSeqLen) + lpad + rpad);
BAM_AlignmentGetSequence(rec, seqDNA + lpad);
if (G.useQUAL) {
uint8_t const *squal;
BAM_AlignmentGetQuality(rec, &squal);
memmove(qual + lpad, squal, readlen);
}
else {
uint8_t const *squal;
uint8_t qoffset = 0;
unsigned i;
rc = BAM_AlignmentGetQuality2(rec, &squal, &qoffset);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "Spot '$(name)': length of original quality does not match sequence", "name=%s", name));
goto LOOP_END;
}
if (qoffset) {
for (i = 0; i != readlen; ++i)
qual[i + lpad] = squal[i] - qoffset;
QUAL_CHANGED_OQ;
}
else
memmove(qual + lpad, squal, readlen);
}
readlen = readlen + lpad + rpad;
data.data.align_group.elements = 0;
data.data.align_group.buffer = alignGroup;
}
if (G.hasTI) {
rc = BAM_AlignmentGetTI(rec, &ti);
if (rc)
ti = 0;
rc = 0;
}
rc = KDataBufferResize(&seqBuffer, readlen);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize record buffer");
goto LOOP_END;
}
rc = KDataBufferResize(&qualBuffer, readlen);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize record buffer");
goto LOOP_END;
}
AR_REF_ORIENT(data) = (flags & BAMFlags_SelfIsReverse) == 0 ? false : true;
rpos = -1;
if (aligned) {
BAM_AlignmentGetPosition(rec, &rpos);
BAM_AlignmentGetRefSeqId(rec, &refSeqId);
if (refSeqId != lastRefSeqId) {
refSeq = NULL;
BAM_FileGetRefSeqById(bam, refSeqId, &refSeq);
}
}
revcmp = (isColorSpace && !aligned) ? false : AR_REF_ORIENT(data);
(void)PLOGMSG(klogDebug, (klogDebug, "Read '$(name)' is $(or) at $(ref):$(pos)", "name=%s,or=%s,ref=%s,pos=%i", name, revcmp ? "reverse" : "forward", refSeq ? refSeq->name : "(none)", rpos));
COPY_READ((INSDC_dna_text*)seqBuffer.base, seqDNA, readlen, revcmp);
COPY_QUAL((uint8_t*)qualBuffer.base, qual, readlen, revcmp);
AR_MAPQ(data) = GetMapQ(rec);
if (!isPrimary && AR_MAPQ(data) < G.minMapQual)
goto LOOP_END;
if (aligned && align == NULL) {
rc = RC(rcApp, rcFile, rcReading, rcData, rcInconsistent);
(void)PLOGERR(klogErr, (klogErr, rc, "File '$(file)' contains aligned records", "file=%s", bamFile));
goto LOOP_END;
}
while (aligned) {
if (rpos >= 0 && refSeqId >= 0) {
if (refSeqId == skipRefSeqID) {
DISCARD_SKIP_REFERENCE;
goto LOOP_END;
}
if (refSeqId == unmapRefSeqId) {
aligned = false;
UNALIGNED_UNALIGNED_REF;
break;
}
unmapRefSeqId = -1;
if (refSeq == NULL) {
rc = SILENT_RC(rcApp, rcFile, rcReading, rcData, rcInconsistent);
(void)PLOGERR(klogWarn, (klogWarn, rc, "File '$(file)': Spot '$(name)' refers to an unknown Reference number $(refSeqId)", "file=%s,refSeqId=%i,name=%s", bamFile, (int)refSeqId, name));
rc = CheckLimitAndLogError();
DISCARD_UNKNOWN_REFERENCE;
goto LOOP_END;
}
else {
bool shouldUnmap = false;
if (G.refFilter && strcmp(G.refFilter, refSeq->name) != 0) {
(void)PLOGMSG(klogInfo, (klogInfo, "Skipping Reference '$(name)'", "name=%s", refSeq->name));
skipRefSeqID = refSeqId;
DISCARD_SKIP_REFERENCE;
goto LOOP_END;
}
/*
if (ctx->references.count(refSeq->name) == 0) {
spdlog::info("Reference: '{}', length: {}", refSeq->name, refSeq->length);
ctx->references.insert(refSeq->name);
}
*/
rc = ReferenceSetFile(ref, refSeq->name, refSeq->length, refSeq->checksum, &shouldUnmap, &wasRenamed);
if (rc == 0) {
lastRefSeqId = refSeqId;
if (shouldUnmap) {
aligned = false;
unmapRefSeqId = refSeqId;
UNALIGNED_UNALIGNED_REF;
}
break;
}
if (GetRCObject(rc) == rcConstraint && GetRCState(rc) == rcViolated) {
int const level = G.limit2config ? klogWarn : klogErr;
(void)PLOGMSG(level, (level, "Could not find a Reference to match { name: '$(name)', length: $(rlen) }", "name=%s,rlen=%u", refSeq->name, (unsigned)refSeq->length));
}
else if (!G.limit2config) {
(void)PLOGERR(klogErr, (klogErr, rc, "File '$(file)': Spot '$(sname)' refers to an unknown Reference '$(rname)'", "file=%s,rname=%s,sname=%s", bamFile, refSeq->name, name));
}
if (G.limit2config) {
rc = 0;
UNALIGNED_UNKNOWN_REF;
}
goto LOOP_END;
}
}
else if (refSeqId < 0) {
(void)PLOGMSG(klogWarn, (klogWarn, "Spot '$(name)' was marked aligned, but reference id = $(id) is invalid", "name=%.*s,id=%i", namelen, name, refSeqId));
if ((rc = CheckLimitAndLogError()) != 0) goto LOOP_END;
UNALIGNED_INVALID_REF;
}
else {
(void)PLOGMSG(klogWarn, (klogWarn, "Spot '$(name)' was marked aligned, but reference position = $(pos) is invalid", "name=%.*s,pos=%i", namelen, name, rpos));
if ((rc = CheckLimitAndLogError()) != 0) goto LOOP_END;
UNALIGNED_INVALID_REF_POS;
}
aligned = false;
}
if (!aligned && (G.refFilter != NULL || G.limit2config)) {
assert(!"this shouldn't happen");
goto LOOP_END;
}
AR_KEY(data) = keyId;
AR_READNO(data) = readNo;
if (wasInserted) {
}
else if (isPrimary || G.assembleWithSecondary || G.deferSecondary) {
/* other times */
int o_pcr_dup = 0;
int const n_pcr_dup = (flags & BAMFlags_IsDuplicate) == 0 ? 0 : 1;
#ifndef NO_METADATA
if (!opt_is_primary.has_value())
opt_is_primary = metadata.get<bit_t>(metadata_t::e_primary_is_set).test(row_id);
if (!opt_is_primary.value()) {
metadata.get<bit_t>(metadata_t::e_primary_is_set).set(row_id);
o_pcr_dup = n_pcr_dup;
opt_is_primary = true;
} else {
if (!opt_pcr_dup.has_value())
opt_pcr_dup = metadata.get<bit_t>(metadata_t::e_pcr_dup).test(row_id);
o_pcr_dup = opt_pcr_dup.value() ? 1 : 0;
}
if (!opt_pcr_dup.has_value() || opt_pcr_dup.value() != (o_pcr_dup & n_pcr_dup)) {
metadata.get<bit_t>(metadata_t::e_pcr_dup).set(row_id, o_pcr_dup & n_pcr_dup);
opt_pcr_dup = o_pcr_dup & n_pcr_dup;
}
#endif
#if defined(HAS_CTX_VALUE)
if (!value->primary_is_set) {
o_pcr_dup = n_pcr_dup;
value->primary_is_set = 1;
} else {
o_pcr_dup = value->pcr_dup;
}
value->pcr_dup = o_pcr_dup & n_pcr_dup;
#endif
if (o_pcr_dup != (o_pcr_dup & n_pcr_dup)) {
FLAG_CHANGED_PCR_DUP;
}
#ifndef NO_METADATA
auto v_unmated = opt_is_unmated.value_or(metadata.get<bit_t>(metadata_t::e_unmated).test(row_id));
#ifdef HAS_CTX_VALUE
if (v_unmated != value->unmated) {
spdlog::error("Inconsistent unmated");
throw runtime_error("Inconsistent unmated");
}
#endif
#else
auto v_unmated = value->unmated;
#endif
if (mated && v_unmated) {
(void)PLOGERR(klogWarn, (klogWarn, SILENT_RC(rcApp, rcFile, rcReading, rcData, rcInconsistent),
"Spot '$(name)', which was first seen without mate info, now has mate info",
"name=%s", name));
rc = CheckLimitAndLogError();
DISCARD_BAD_FRAGMENT_INFO;
goto LOOP_END;
}
else if (!mated && !v_unmated) {
(void)PLOGERR(klogWarn, (klogWarn, SILENT_RC(rcApp, rcFile, rcReading, rcData, rcInconsistent),
"Spot '$(name)', which was first seen with mate info, now has no mate info",
"name=%s", name));
rc = CheckLimitAndLogError();
DISCARD_BAD_FRAGMENT_INFO;
goto LOOP_END;
}
}
if (isPrimary) {
switch (readNo) {
case 1: {
#if !defined NO_METADATA
if (!primaryId[0])
primaryId[0] = wasInserted ? 0 : metadata.get<u64_t>(metadata_t::E_PRIM_ID[0]).get_no_check(row_id);
auto v = primaryId[0];
#ifdef HAS_CTX_VALUE
if (v != CTX_VALUE_GET_P_ID(*value, 0)) {
spdlog::error("Inconsistent primaryId");
throw runtime_error("Inconsistent primaryId");
}
#endif
#else
auto v = CTX_VALUE_GET_P_ID(*value, 0);
#endif
if (v != 0) {
isPrimary = false;
FLAG_CHANGED_PRIMARY_DUP;
}
else if (aligned) {
#if !defined NO_METADATA
auto v_unaligned_1 = wasInserted ? 0 : metadata.get<bit_t>(metadata_t::e_unaligned_1).test(row_id);
#ifdef HAS_CTX_VALUE
if (v_unaligned_1 != value->unaligned_1) {
spdlog::error("Inconsistent v_unaligned_1");
throw runtime_error("Inconsistent v_unaligned_1");
}
#endif
#else
auto v_unaligned_1 = value->unaligned_1;
#endif
if (v_unaligned_1) {
(void)PLOGMSG(klogWarn, (klogWarn, "Read 1 of spot '$(name)', which was unmapped, is now being mapped at position $(pos) on reference '$(ref)'; this alignment will be considered as secondary", "name=%s,ref=%s,pos=%u", name, refSeq->name, rpos));
isPrimary = false;
FLAG_CHANGED_WAS_UNALIGNED;
}
}
break;
}
case 2:
{
#ifndef NO_METADATA
if (!primaryId[1])
primaryId[1] = wasInserted ? 0 : metadata.get<u64_t>(metadata_t::E_PRIM_ID[1]).get_no_check(row_id);
auto v = primaryId[1];
#ifdef HAS_CTX_VALUE
if (v != CTX_VALUE_GET_P_ID(*value, 1)) {
spdlog::error("Inconsistent primaryId[1]");
throw runtime_error("Inconsistent primaryId[1]");
}
#endif
#else
auto v = CTX_VALUE_GET_P_ID(*value, 1);
#endif
if (v != 0) {
isPrimary = false;
FLAG_CHANGED_PRIMARY_DUP;
}
else if (aligned) {
#ifndef NO_METADATA
auto v_unaligned_2 = wasInserted ? 0 : metadata.get<bit_t>(metadata_t::e_unaligned_2).test(row_id);
#ifdef HAS_CTX_VALUE
if (v_unaligned_2 != value->unaligned_2) {
spdlog::error("Inconsistent v_unaligned_2");
throw runtime_error("Inconsistent v_unaligned_2");
}
#endif
#else
auto v_unaligned_2 = value->unaligned_2;
#endif
if (v_unaligned_2) {
(void)PLOGMSG(klogWarn, (klogWarn, "Read 2 of spot '$(name)', which was unmapped, is now being mapped at position $(pos) on reference '$(ref)'; this alignment will be considered as secondary", "name=%s,ref=%s,pos=%u", name, refSeq->name, rpos));
isPrimary = false;
FLAG_CHANGED_WAS_UNALIGNED;
}
}
break;
}
default:
break;
}
}
if (hardclipped) {
#ifndef NO_METADATA
metadata.get<bit_t>(metadata_t::e_hardclipped).set(row_id);
#endif
#if defined(HAS_CTX_VALUE)
value->hardclipped = 1;
#endif
}
#if 0 /** EY TO REVIEW **/
if (!isPrimary && value->hardclipped) {
DISCARD_HARDCLIP_SECONDARY;
goto LOOP_END;
}
#endif
/* input is clean */
++recordsProcessed;
data.isPrimary = isPrimary;
if (aligned) {
uint32_t matches = 0;
uint32_t misses = 0;
uint8_t rna_orient = ' ';
FixOverhangingAlignment(&cigBuf, &opCount, rpos, refSeq->length, readlen);
BAM_AlignmentGetRNAStrand(rec, &rna_orient);
{
int const intronType = rna_orient == '+' ? NCBI_align_ro_intron_plus :
rna_orient == '-' ? NCBI_align_ro_intron_minus :
hasCG ? NCBI_align_ro_complete_genomics :
NCBI_align_ro_intron_unknown;
rc = ReferenceRead(ref, &data, rpos, (const uint32_t*)cigBuf.base, opCount, seqDNA, readlen, intronType, &matches, &misses);
}
if (rc == 0) {
int const i = readNo - 1;
int const clipped_rl = readlen < 255 ? readlen : 255;
if (i >= 0 && i < 2) {
#if !defined NO_METADATA
int const rl = wasInserted ? 0 : opt_frag_len[i].value_or(metadata.get<u16_t>(metadata_t::E_FRAG_LEN[i]).get_no_check(row_id));
#ifdef HAS_CTX_VALUE
if (rl != value->fragment_len[i]) {
spdlog::error("Inconsistent fragment_len");
throw runtime_error("Inconsistent fragment_len");
}
#endif
#else
int const rl = value->fragment_len[i];
#endif
if (rl == 0) {
#if !defined NO_METADATA
metadata.get<u16_t>(metadata_t::E_FRAG_LEN[i]).set(row_id, clipped_rl);
#endif
#ifdef HAS_CTX_VALUE
value->fragment_len[i] = clipped_rl;
#endif
}
else if (rl != clipped_rl) {
if (isPrimary) {
rc = RC(rcApp, rcFile, rcReading, rcConstraint, rcViolated);
(void)PLOGERR(klogErr, (klogErr, rc, "Primary alignment for '$(name)' has different length ($(len)) than previously recorded secondary alignment. Try to defer secondary alignment processing.",
"name=%s,len=%d", name, readlen));
}
else {
rc = SILENT_RC(rcApp, rcFile, rcReading, rcConstraint, rcViolated);
(void)PLOGERR(klogWarn, (klogWarn, rc, "Secondary alignment for '$(name)' has different length ($(len)) than previously recorded primary alignment; discarding secondary alignment.",
"name=%s,len=%d", name, readlen));
DISCARD_BAD_SECONDARY;
rc = CheckLimitAndLogError();
}
goto LOOP_END;
}
}
}
if (rc == 0 && (matches < G.minMatchCount || (matches == 0 && !G.acceptNoMatch))) {
if (isPrimary) {
if (misses > matches) {
RecordNoMatch(name, refSeq->name, rpos);
rc = LogNoMatch(name, refSeq->name, (unsigned)rpos, (unsigned)matches);
if (rc)
goto LOOP_END;
}
}
else {
(void)PLOGMSG(klogWarn, (klogWarn, "Spot '$(name)' contains too few ($(count)) matching bases to reference '$(ref)' at $(pos); discarding secondary alignment",
"name=%s,ref=%s,pos=%u,count=%u", name, refSeq->name, (unsigned)rpos, (unsigned)matches));
DISCARD_BAD_SECONDARY;
rc = 0;
goto LOOP_END;
}
}
if (rc) {
aligned = false;
if (((int)GetRCObject(rc)) == ((int)rcData) && GetRCState(rc) == rcNotAvailable) {
/* because of code above converting hard clips to soft clips, this should be unreachable */
abort();
}
else if (((int)GetRCObject(rc)) == ((int)rcData)) {
UNALIGNED_INVALID_INFO;
(void)PLOGERR(klogWarn, (klogWarn, rc, "Spot '$(name)': bad alignment to reference '$(ref)' at $(pos)", "name=%s,ref=%s,pos=%u", name, refSeq->name, rpos));
/* Data errors may get reset; alignment will be unmapped at any rate */
rc = CheckLimitAndLogError();
}
else {
UNALIGNED_INVALID_REF_POS;
(void)PLOGERR(klogWarn, (klogWarn, rc, "Spot '$(name)': error reading reference '$(ref)' at $(pos)", "name=%s,ref=%s,pos=%u", name, refSeq->name, rpos));
rc = CheckLimitAndLogError();
}
if (rc) goto LOOP_END;
}
}
if (!aligned && isPrimary) {
switch (readNo) {
case 1:
#if !defined NO_METADATA
metadata.get<bit_t>(metadata_t::e_unaligned_1).set(row_id);
#endif
#ifdef HAS_CTX_VALUE
value->unaligned_1 = 1;
#endif
break;
case 2:
#if !defined NO_METADATA
metadata.get<bit_t>(metadata_t::e_unaligned_2).set(row_id);
#endif
#ifdef HAS_CTX_VALUE
value->unaligned_2 = 1;
#endif
break;
default:
break;
}
}
if (isPrimary && aligned) {
#if !defined NO_METADATA
if (!primaryId[readNo-1])
primaryId[readNo-1] = wasInserted ? 0 : metadata.get<u64_t>(metadata_t::E_PRIM_ID[readNo-1]).get_no_check(row_id);
auto v = primaryId[readNo-1];
#ifdef HAS_CTX_VALUE
if (v != CTX_VALUE_GET_P_ID(*value, readNo - 1)) {
spdlog::error("Inconsistent CTX_VALUE_GET_P_ID");
throw runtime_error("Inconsistent CTX_VALUE_GET_P_ID");
}
#endif
#else
auto v = CTX_VALUE_GET_P_ID(*value, readNo - 1);
#endif
switch (readNo) {
case 1:
if (v == 0) {
data.alignId = ++ctx->primaryId;
#ifndef NO_METADATA
metadata.get<u64_t>(metadata_t::E_PRIM_ID[0]).set(row_id, data.alignId);
#endif
#if defined(HAS_CTX_VALUE)
CTX_VALUE_SET_P_ID(*value, 0, data.alignId);
#endif
}
break;
case 2:
if (v == 0) {
data.alignId = ++ctx->primaryId;
#ifndef NO_METADATA
metadata.get<u64_t>(metadata_t::E_PRIM_ID[1]).set(row_id, data.alignId);
#endif
#if defined(HAS_CTX_VALUE)
CTX_VALUE_SET_P_ID(*value, 1, data.alignId);
#endif
}
break;
default:
break;
}
}
if (G.mode == mode_Archive)
goto WRITE_SEQUENCE;
else
goto WRITE_ALIGNMENT;
if (0) {
WRITE_SEQUENCE:
#ifndef NO_METADATA
// int64_t const spotId = metadata.Uint64(e_spotId).get_no_check(row_id);
int64_t const spotId = wasInserted ? 0 : metadata.get<u64_t>(metadata_t::e_spotId).get_no_check(row_id);
#ifdef HAS_CTX_VALUE
if (spotId != CTX_VALUE_GET_S_ID(*value)) {
spdlog::error("Inconsistent spotId");
throw runtime_error("Inconsistent spotId");
}
#endif
#else
int64_t const spotId = CTX_VALUE_GET_S_ID(*value);
#endif
if (mated) {
bool const spotHasBeenWritten = (spotId != 0);
if (spotHasBeenWritten == false) {
#ifndef NO_METADATA
uint32_t fragmentId = wasInserted ? 0 : metadata.get<u32_t>(metadata_t::e_fragmentId).get_no_check(row_id);
#ifdef HAS_CTX_VALUE
if (value->fragmentId != fragmentId) {
spdlog::error("Inconsistent fragmentId");
throw runtime_error("Inconsistent fragmentId");
}
#endif
#else
uint32_t fragmentId = value->fragmentId;
#endif
bool const spotHasFragmentInfo = (fragmentId != 0);
bool const spotIsFirstSeen = spotHasFragmentInfo ? false : true;
if (spotIsFirstSeen) {
if (!isPrimary) {
if ( (!G.assembleWithSecondary || hardclipped) && !G.deferSecondary ) {
goto WRITE_ALIGNMENT;
}
(void)PLOGMSG(klogDebug, (klogDebug, "Spot '$(name)' (id $(id)) is being constructed from secondary alignment information", "id=%lx,name=%s", keyId, name));
}
/* start spot assembly */
unsigned sz;
FragmentInfo fi;
int32_t mate_refSeqId = -1;
int64_t pnext = 0;
if (ctx->m_isSingleGroup == false) // otherwise spotGroup was captured
getSpotGroup(rec, spotGroup);
BAM_AlignmentGetBarCode(rec, &barCode);
if (barCode) {
if (spotGroup[0] != '\0' && rec->platform == SRA_PLATFORM_UNDEFINED) {
/* don't use bar code */
}
else {
unsigned const sglen = strlen(barCode);
if (sglen + 1 < sizeof(spotGroup))
memmove(spotGroup, barCode, sglen + 1);
}
}
memset(&fi, 0, sizeof(fi));
fi.aligned = isPrimary ? aligned : 0;
fi.ti = ti;
fi.orientation = AR_REF_ORIENT(data);
fi.readNo = readNo;
fi.sglen = strlen(spotGroup);
fi.lglen = strlen(linkageGroup);
fi.readlen = readlen;
fi.cskey = cskey;
fi.is_bad = (flags & BAMFlags_IsLowQuality) != 0;
sz = sizeof(fi) + 2*fi.readlen + fi.sglen + fi.lglen;
if (align) {
BAM_AlignmentGetMateRefSeqId(rec, &mate_refSeqId);
BAM_AlignmentGetMatePosition(rec, &pnext);
}
if(align && mate_refSeqId == refSeqId && pnext > 0 && pnext!=rpos /*** weird case in some bams**/){
rc = MemBankAlloc(ctx->frags, &fragmentId, sz, 0, false);
fcountBoth++;
} else {
rc = MemBankAlloc(ctx->frags, &fragmentId, sz, 0, true);
fcountOne++;
}
#ifndef NO_METADATA
metadata.get<u32_t>(metadata_t::e_fragmentId).set(row_id, fragmentId);
#endif
#if defined (HAS_CTX_VALUE)
value->fragmentId = fragmentId;
#endif
if (rc) {
(void)LOGERR(klogErr, rc, "KMemBankAlloc failed");
goto LOOP_END;
}
/*printf("IN:%10d\tcnt2=%ld\tcnt1=%ld\n",value->fragmentId,fcountBoth,fcountOne);*/
rc = KDataBufferResize(&fragBuf, sz);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize fragment buffer");
goto LOOP_END;
}
{{
uint8_t *dst = (uint8_t*) fragBuf.base;
memmove(dst,&fi,sizeof(fi));
dst += sizeof(fi);
memmove(dst, seqBuffer.base, readlen);
dst += readlen;
memmove(dst, qualBuffer.base, readlen);
dst += fi.readlen;
memmove(dst, spotGroup, fi.sglen);
dst += fi.sglen;
memmove(dst, linkageGroup, fi.lglen);
dst += fi.lglen;
}}
rc = MemBankWrite(ctx->frags, fragmentId, 0, fragBuf.base, sz, &rsize);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "KMemBankWrite failed writing fragment $(id)", "id=%u", fragmentId));
goto LOOP_END;
}
if (revcmp) {
QUAL_CHANGED_REVERSED;
SEQ__CHANGED_REV_COMP;
}
}
else if (spotHasFragmentInfo) {
/* continue spot assembly */
FragmentInfo *fip;
{
size_t size1;
size_t size2;
rc = MemBankSize(ctx->frags, fragmentId, &size1);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "KMemBankSize failed on fragment $(id)", "id=%u", fragmentId));
goto LOOP_END;
}
rc = KDataBufferResize(&fragBuf, size1);
fip = (FragmentInfo *)fragBuf.base;
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "Failed to resize fragment buffer", ""));
goto LOOP_END;
}
rc = MemBankRead(ctx->frags, fragmentId, 0, fragBuf.base, size1, &size2);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "KMemBankRead failed on fragment $(id)", "id=%u", fragmentId));
goto LOOP_END;
}
assert(size1 == size2);
}
if (readNo == fip->readNo) {
/* is a repeat of the same read; do nothing */
}
else {
/* mate found; finish spot assembly */
unsigned read1 = 0;
unsigned read2 = 1;
char const *const seq1 = (const char *)&fip[1];
char const *const qual1 = (const char *)(seq1 + fip->readlen);
char const *const sg1 = (const char *)(qual1 + fip->readlen);
char const *const bx1 = (const char *)(sg1 + fip->sglen);
if (!isPrimary) {
if ((!G.assembleWithSecondary || hardclipped) && !G.deferSecondary ) {
goto WRITE_ALIGNMENT;
}
(void)PLOGMSG(klogDebug, (klogDebug, "Spot '$(name)' (id $(id)) is being constructed from secondary alignment information", "id=%lx,name=%s", keyId, name));
}
rc = KDataBufferResize(&seqBuffer, readlen + fip->readlen);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize record buffer");
goto LOOP_END;
}
rc = KDataBufferResize(&qualBuffer, readlen + fip->readlen);
if (rc) {
(void)LOGERR(klogErr, rc, "Failed to resize record buffer");
goto LOOP_END;
}
if (readNo < fip->readNo) {
read1 = 1;
read2 = 0;
}
memset(&srecStorage, 0, sizeof(srecStorage));
srec.numreads = 2;
srec.readLen[read1] = fip->readlen;
srec.readLen[read2] = readlen;
srec.readStart[1] = srec.readLen[0];
{
char const *const s1 = seq1;
char const *const s2 = (const char*)seqBuffer.base;
char *const d = (char*)seqBuffer.base;
char *const d1 = d + srec.readStart[read1];
char *const d2 = d + srec.readStart[read2];
srec.seq = (char*)seqBuffer.base;
if (d2 != s2) {
memmove(d2, s2, readlen);
}
memmove(d1, s1, fip->readlen);
}
{
char const *const s1 = qual1;
char const *const s2 = (const char*)qualBuffer.base;
char *const d = (char*)qualBuffer.base;
char *const d1 = d + srec.readStart[read1];
char *const d2 = d + srec.readStart[read2];
srec.qual = (uint8_t*)qualBuffer.base;
if (d2 != s2) {
memmove(d2, s2, readlen);
}
memmove(d1, s1, fip->readlen);
}
srec.ti[read1] = fip->ti;
srec.ti[read2] = ti;
srec.aligned[read1] = fip->aligned;
srec.aligned[read2] = isPrimary ? aligned : 0;
srec.is_bad[read1] = fip->is_bad;
srec.is_bad[read2] = (flags & BAMFlags_IsLowQuality) != 0;
srec.orientation[read1] = fip->orientation;
srec.orientation[read2] = AR_REF_ORIENT(data);
srec.cskey[read1] = fip->cskey;
srec.cskey[read2] = cskey;
srec.keyId = keyId;
srec.spotGroup = sg1;
srec.spotGroupLen = fip->sglen;
srec.linkageGroup = bx1;
srec.linkageGroupLen = fip->lglen;
srec.seq = (char*)seqBuffer.base;
srec.qual = (uint8_t*)qualBuffer.base;
#ifndef NO_METADATA
auto v_pcr_dup = opt_pcr_dup.value_or(metadata.get<bit_t>(metadata_t::e_pcr_dup).test(row_id));
#ifdef HAS_CTX_VALUE
if (v_pcr_dup != value->pcr_dup) {
spdlog::error("Inconsistent pcr_dup");
throw runtime_error("Inconsistent pcr_dup");
}
#endif
#else
auto v_pcr_dup = value->pcr_dup;
#endif
rc = SequenceWriteRecord(seq, &srec, isColorSpace, v_pcr_dup, rec->platform);
if (rc) {
(void)LOGERR(klogErr, rc, "SequenceWriteRecord failed");
goto LOOP_END;
}
++ctx->spotId;
#ifndef NO_METADATA
//*ctx->m_fs << "mate:" << keyId << '\t' << ctx->spotId << '\t' << row_id << endl;
//ctx->key4spot[ctx->spotId] = keyId;
metadata.get<u64_t>(metadata_t::e_spotId).set(row_id, ctx->spotId);
#endif
#if defined (HAS_CTX_VALUE)
CTX_VALUE_SET_S_ID(*value, ctx->spotId);
#endif
if(fragmentId & 1){
fcountOne--;
} else {
fcountBoth--;
}
/* printf("OUT:%9d\tcnt2=%ld\tcnt1=%ld\n",fragmentId,fcountBoth,fcountOne);*/
rc = MemBankFree(ctx->frags, fragmentId);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "KMemBankFree failed on fragment $(id)", "id=%u", fragmentId));
goto LOOP_END;
}
#ifndef NO_METADATA
//fragment_buffer.set_bit_no_check(row_id);
metadata.get<u32_t>(metadata_t::e_fragmentId).set(row_id, 0);
#endif
#if defined (HAS_CTX_VALUE)
value->fragmentId = 0;
#endif
if (revcmp) {
QUAL_CHANGED_REVERSED;
SEQ__CHANGED_REV_COMP;
}
if (v_pcr_dup && (srec.is_bad[0] || srec.is_bad[1])) {
FLAG_CHANGED_400_AND_200;
filterFlagConflictRecords++;
if (filterFlagConflictRecords < MAX_WARNINGS_FLAG_CONFLICT) {
(void)PLOGMSG(klogWarn, (klogWarn, "Spot '$(name)': both 0x400 and 0x200 flag bits set, only 0x400 will be saved", "name=%s", name));
}
else if (filterFlagConflictRecords == MAX_WARNINGS_FLAG_CONFLICT) {
(void)PLOGMSG(klogWarn, (klogWarn, "Last reported warning: Spot '$(name)': both 0x400 and 0x200 flag bits set, only 0x400 will be saved", "name=%s", name));
}
}
}
}
else {
(void)PLOGMSG(klogErr, (klogErr, "Spot '$(name)' has caused the loader to enter an illogical state", "name=%s", name));
assert("this should never happen");
}
} else { // Spot has been written
}
}
else if (spotId == 0) {
/* new unmated fragment - no spot assembly */
if (!isPrimary) {
if ((!G.assembleWithSecondary || hardclipped) && !G.deferSecondary ) {
goto WRITE_ALIGNMENT;
}
(void)PLOGMSG(klogDebug, (klogDebug, "Spot '$(name)' (id $(id)) is being constructed from secondary alignment information", "id=%lx,name=%s", keyId, name));
}
if (ctx->m_isSingleGroup == false) // otherwise spotGroup was captured
getSpotGroup(rec, spotGroup);
BAM_AlignmentGetBarCode(rec, &barCode);
if (barCode) {
if (spotGroup[0] != '\0' && rec->platform == SRA_PLATFORM_UNDEFINED) {
/* don't use bar code */
}
else {
unsigned const sglen = strlen(barCode);
if (sglen + 1 < sizeof(spotGroup))
memmove(spotGroup, barCode, sglen + 1);
}
}
memset(&srecStorage, 0, sizeof(srecStorage));
srec.numreads = 1;
srec.readLen[0] = readlen;
srec.ti[0] = ti;
srec.aligned[0] = isPrimary ? aligned : 0;
srec.is_bad[0] = (flags & BAMFlags_IsLowQuality) != 0;
srec.orientation[0] = AR_REF_ORIENT(data);
srec.cskey[0] = cskey;
srec.keyId = keyId;
srec.spotGroup = spotGroup;
srec.spotGroupLen = strlen(spotGroup);
srec.linkageGroup = linkageGroup;
srec.linkageGroupLen = strlen(linkageGroup);
srec.seq = (char*)seqBuffer.base;
srec.qual = (uint8_t*)qualBuffer.base;
#ifndef NO_METADATA
auto v_pcr_dup = opt_pcr_dup.value_or(metadata.get<bit_t>(metadata_t::e_pcr_dup).test(row_id));
#ifdef HAS_CTX_VALUE
if (v_pcr_dup != value->pcr_dup) {
spdlog::error("Inconsistent pcr_dup");
throw runtime_error("Inconsistent pcr_dup");
}
#endif
#else
auto v_pcr_dup = value->pcr_dup;
#endif
rc = SequenceWriteRecord(seq, &srec, isColorSpace, v_pcr_dup, rec->platform);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "SequenceWriteRecord failed", ""));
goto LOOP_END;
}
++ctx->spotId;
#ifndef NO_METADATA
// *ctx->m_fs << "no mate:" << keyId << '\t' << ctx->spotId << '\t' << row_id << endl;
// ctx->key4spot[ctx->spotId] = keyId;
metadata.get<u64_t>(metadata_t::e_spotId).set(row_id, ctx->spotId);
//fragment_buffer.set_bit_no_check(row_id);
metadata.get<u32_t>(metadata_t::e_fragmentId).set(row_id, 0);
#endif
#if defined (HAS_CTX_VALUE)
CTX_VALUE_SET_S_ID(*value, ctx->spotId);
value->fragmentId = 0;
#endif
if (v_pcr_dup && srec.is_bad[0]) {
FLAG_CHANGED_400_AND_200;
filterFlagConflictRecords++;
if (filterFlagConflictRecords < MAX_WARNINGS_FLAG_CONFLICT) {
(void)PLOGMSG(klogWarn, (klogWarn, "Spot '$(name)': both 0x400 and 0x200 flag bits set, only 0x400 will be saved", "name=%s", name));
}
else if (filterFlagConflictRecords == MAX_WARNINGS_FLAG_CONFLICT) {
(void)PLOGMSG(klogWarn, (klogWarn, "Last reported warning: Spot '$(name)': both 0x400 and 0x200 flag bits set, only 0x400 will be saved", "name=%s", name));
}
}
if (revcmp) {
QUAL_CHANGED_REVERSED;
SEQ__CHANGED_REV_COMP;
}
}
}
WRITE_ALIGNMENT:
if (aligned) {
if (mated && !isPrimary) {
int32_t bam_mrid;
int64_t mpos;
int64_t mrid = 0;
int64_t tlen;
BAM_AlignmentGetMatePosition(rec, &mpos);
BAM_AlignmentGetMateRefSeqId(rec, &bam_mrid);
BAM_AlignmentGetInsertSize(rec, &tlen);
if (mpos >= 0 && bam_mrid >= 0 && tlen != 0) {
BAMRefSeq const *mref;
BAM_FileGetRefSeq(bam, bam_mrid, &mref);
if (mref) {
rc_t rc_temp = ReferenceGet1stRow(ref, &mrid, mref->name);
if (rc_temp == 0) {
data.mate_ref_pos = mpos;
data.template_len = tlen;
data.mate_ref_orientation = (flags & BAMFlags_MateIsReverse) ? 1 : 0;
}
else {
(void)PLOGERR(klogWarn, (klogWarn, rc_temp, "Failed to get refID for $(name)", "name=%s", mref->name));
MATE_INFO_LOST_UNKNOWN_REF;
}
data.mate_ref_id = mrid;
}
else {
MATE_INFO_LOST_INVALID;
}
}
else if (mpos >= 0 || bam_mrid >= 0 || tlen != 0) {
MATE_INFO_LOST_MISSING;
}
}
if (wasRenamed) {
RENAMED_REFERENCE;
}
#ifndef NO_METADATA
auto v_aln_count = wasInserted ? 0 : metadata.get<u16_t>(metadata_t::E_ALN_COUNT[readNo - 1]).get_no_check(row_id);
#ifdef HAS_CTX_VALUE
if (v_aln_count != value->alignmentCount[readNo - 1]) {
spdlog::error("Inconsistent alignmentCount");
throw runtime_error("Inconsistent alignmentCount");
}
#endif
if (v_aln_count < 254) {
#ifdef HAS_CTX_VALUE
++value->alignmentCount[readNo - 1];
#endif
metadata.get<u16_t>(metadata_t::E_ALN_COUNT[readNo - 1]).inc(row_id);
}
#else
auto v_aln_count = value->alignmentCount[readNo - 1];
if (v_aln_count < 254) {
++value->alignmentCount[readNo - 1];
}
#endif
++ctx->alignCount;
if (linkageGroup[0] != '\0') {
AR_LINKAGE_GROUP(data).elements = strlen(linkageGroup);
AR_LINKAGE_GROUP(data).buffer = linkageGroup;
}
rc = AlignmentWriteRecord(align, &data);
if (rc == 0) {
if (!isPrimary)
data.alignId = ++ctx->secondId;
rc = ReferenceAddAlignId(ref, data.alignId, isPrimary);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "ReferenceAddAlignId failed", ""));
}
else {
*had_alignments = true;
}
}
else {
(void)PLOGERR(klogErr, (klogErr, rc, "AlignmentWriteRecord failed", ""));
}
}
/**************************************************************/
LOOP_END:
BAM_AlignmentRelease(rec);
#if !defined(NEW_QUEUE)
delete queue_rec;
#endif
++reccount;
if (G.maxAlignCount > 0 && reccount >= G.maxAlignCount)
break;
if (rc == 0)
*had_sequences = true;
else
break;
}
spdlog::info("New spots: {:L}, recordRead: {:L}, recordsProcess: {:L}", new_spots, recordsRead, recordsProcessed);
/*
for (const auto& it : ctx->spots) {
spdlog::info("Orhpan spot: {}", it);
}
*/
//ctx->m_fs->close();
#ifndef NEW_QUEUE
if (bamread_thread != NULL && bamq != NULL) {
KQueueSeal(bamq);
for ( ; ; ) {
timeout_t tm;
void *rr = NULL;
rc_t rc2;
TimeoutInit(&tm, 1000);
rc2 = KQueuePop(bamq, &rr, &tm);
if (rc2) break;
BAM_AlignmentRelease((BAM_Alignment *)rr);
}
KThreadWait(bamread_thread, NULL);
}
#else
rw_done.store(true);
KThreadWait(bamread_thread, NULL);
{
queue_rec_t queue_rec;
while (rw_queue.try_dequeue(queue_rec)) {
//spdlog::info("There still recs, dude!");
BAM_AlignmentRelease(queue_rec.alignment);
}
}
#endif
KThreadRelease(bamread_thread);
#ifndef NEW_QUEUE
KQueueRelease(bamq);
#endif
if (rc) {
if ( (GetRCModule(rc) == rcCont && (int)GetRCObject(rc) == rcData && GetRCState(rc) == rcDone)
|| (GetRCModule(rc) == rcAlign && GetRCObject(rc) == rcRow && GetRCState(rc) == rcNotFound))
{
(void)PLOGMSG(klogInfo, (klogInfo, "EOF '$(file)'; processed $(proc)", "file=%s,read=%lu,proc=%lu", bamFile, (unsigned long)recordsRead, (unsigned long)recordsProcessed));
rc = 0;
}
else {
(void)PLOGERR(klogInfo, (klogInfo, rc, "Error '$(file)'; read $(read); processed $(proc)", "file=%s,read=%lu,proc=%lu", bamFile, (unsigned long)recordsRead, (unsigned long)recordsProcessed));
}
}
if (filterFlagConflictRecords > 0) {
(void)PLOGMSG(klogWarn, (klogWarn, "$(cnt1) out of $(cnt2) records contained warning : both 0x400 and 0x200 flag bits set, only 0x400 will be saved", "cnt1=%lu,cnt2=%lu", filterFlagConflictRecords,recordsProcessed));
}
if (rc == 0 && recordsProcessed == 0) {
(void)LOGMSG(klogWarn, (G.limit2config || G.refFilter != NULL) ?
"All records from the file were filtered out" :
"The file contained no records that were processed.");
rc = RC(rcAlign, rcFile, rcReading, rcData, rcEmpty);
}
BAM_FileRelease(bam);
#ifdef HAS_CTX_VALUE
MMArrayLock(ctx->id2value);
#endif
KDataBufferWhack(&seqBuffer);
KDataBufferWhack(&qualBuffer);
KDataBufferWhack(&buf);
KDataBufferWhack(&fragBuf);
KDataBufferWhack(&cigBuf);
KDataBufferWhack(&data.buffer);
return rc;
}
#if defined(HAS_CTX_VALUE) && defined(NO_METADATA)
static rc_t WriteSoloFragments(context_t *ctx, Sequence *seq)
{
uint32_t i;
unsigned j;
uint64_t idCount = 0;
rc_t rc;
KDataBuffer fragBuf;
SequenceRecordStorage srecStorage;
SequenceRecord srec;
++ctx->pass;
memset(&srec, 0, sizeof(srec));
srec.ti = srecStorage.ti;
srec.readStart = srecStorage.readStart;
srec.readLen = srecStorage.readLen;
srec.orientation = srecStorage.orientation;
srec.is_bad = srecStorage.is_bad;
srec.alignmentCount = srecStorage.alignmentCount;
srec.aligned = srecStorage.aligned;
srec.cskey = srecStorage. cskey;
rc = KDataBufferMake(&fragBuf, 8, 0);
if (rc) {
(void)LOGERR(klogErr, rc, "KDataBufferMake failed");
return rc;
}
// for (idCount = 0, j = 0; j < ctx->keyToID.key2id_count; ++j) {
// idCount += ctx->keyToID.idCount[j];
// }
// KLoadProgressbar_Append(ctx->progress[ctx->pass - 1], idCount);
ctx->visit_keyId([&](uint64_t keyId) {
ctx_value_t *value;
size_t rsize;
size_t sz;
char const *src;
FragmentInfo const *fip;
rc = MMArrayGet(ctx->id2value, (void **)&value, keyId);
if (rc)
return;
KLoadProgressbar_Process(ctx->progress[ctx->pass - 1], 1, false);
if (value->fragmentId == 0)
return;
rc = MemBankSize(ctx->frags, value->fragmentId, &sz);
if (rc) {
(void)LOGERR(klogErr, rc, "KMemBankSize failed");
return;
}
rc = KDataBufferResize(&fragBuf, (size_t)sz);
if (rc) {
(void)LOGERR(klogErr, rc, "KDataBufferResize failed");
return;
}
rc = MemBankRead(ctx->frags, value->fragmentId, 0, fragBuf.base, sz, &rsize);
if (rc) {
(void)LOGERR(klogErr, rc, "KMemBankRead failed");
return;
}
assert( rsize == sz );
fip = (const FragmentInfo*)fragBuf.base;
src = (char const *)&fip[1];
memset(&srecStorage, 0, sizeof(srecStorage));
if (value->unmated) {
srec.numreads = 1;
srec.readLen[0] = fip->readlen;
srec.ti[0] = fip->ti;
srec.aligned[0] = fip->aligned;
srec.is_bad[0] = fip->is_bad;
srec.orientation[0] = fip->orientation;
srec.cskey[0] = fip->cskey;
}
else {
unsigned const read = ((fip->aligned && CTX_VALUE_GET_P_ID(*value, 0) == 0) || value->unaligned_2) ? 1 : 0;
srec.numreads = 2;
srec.readLen[read] = fip->readlen;
srec.readStart[1] = srec.readLen[0];
srec.ti[read] = fip->ti;
srec.aligned[read] = fip->aligned;
srec.is_bad[read] = fip->is_bad;
srec.orientation[read] = fip->orientation;
srec.cskey[0] = srec.cskey[1] = 'N';
srec.cskey[read] = fip->cskey;
}
srec.seq = (char *)src;
srec.qual = (uint8_t *)(src + fip->readlen);
srec.spotGroup = (char *)(src + 2 * fip->readlen);
srec.spotGroupLen = fip->sglen;
srec.linkageGroup = (char *)(src + 2 * fip->readlen * fip->sglen);
srec.linkageGroupLen = fip->lglen;
srec.keyId = keyId;
assert(false);
rc = SequenceWriteRecord(seq, &srec, ctx->isColorSpace, value->pcr_dup, value->platform);
if (rc) {
(void)LOGERR(klogErr, rc, "SequenceWriteRecord failed");
return;
}
/*rc = KMemBankFree(frags, id);*/
CTX_VALUE_SET_S_ID(*value, ++ctx->spotId);
});
MMArrayLock(ctx->id2value);
KDataBufferWhack(&fragBuf);
return rc;
}
static rc_t SequenceUpdateAlignInfo(context_t *ctx, Sequence *seq)
{
rc_t rc = 0;
uint64_t row;
uint64_t keyId;
++ctx->pass;
KLoadProgressbar_Append(ctx->progress[ctx->pass - 1], ctx->spotId + 1);
for (row = 1; row <= ctx->spotId; ++row) {
ctx_value_t *value;
rc = SequenceReadKey(seq, row, &keyId);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "Failed to get key for row $(row)", "row=%u", (unsigned)row));
break;
}
rc = MMArrayGet(ctx->id2value, (void **)&value, keyId);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "Failed to read info for row $(row), index $(idx)", "row=%u,idx=%u", (unsigned)row, (unsigned)keyId));
break;
}
if (G.mode == mode_Remap) {
CTX_VALUE_SET_S_ID(*value, row);
}
if (row != CTX_VALUE_GET_S_ID(*value)) {
rc = RC(rcApp, rcTable, rcWriting, rcData, rcUnexpected);
(void)PLOGMSG(klogErr, (klogErr, "Unexpected spot id $(spotId) for row $(row), index $(idx)", "spotId=%u,row=%u,idx=%u", (unsigned)CTX_VALUE_GET_S_ID(*value), (unsigned)row, (unsigned)keyId));
break;
}
{{
int64_t primaryId[2];
int const logLevel = klogWarn; /*G.assembleWithSecondary ? klogWarn : klogErr;*/
primaryId[0] = CTX_VALUE_GET_P_ID(*value, 0);
primaryId[1] = CTX_VALUE_GET_P_ID(*value, 1);
if (primaryId[0] == 0 && value->alignmentCount[0] != 0) {
rc = RC(rcApp, rcTable, rcWriting, rcConstraint, rcViolated);
(void)PLOGERR(logLevel, (logLevel, rc, "Spot id $(id) read 1 never had a primary alignment", "id=%lx", keyId));
}
if (!value->unmated && primaryId[1] == 0 && value->alignmentCount[1] != 0) {
rc = RC(rcApp, rcTable, rcWriting, rcConstraint, rcViolated);
(void)PLOGERR(logLevel, (logLevel, rc, "Spot id $(id) read 2 never had a primary alignment", "id=%lx", keyId));
}
if (rc != 0 && logLevel == klogErr)
break;
rc = SequenceUpdateAlignData(seq, row, value->unmated ? 1 : 2,
primaryId,
value->alignmentCount);
}}
if (rc) {
(void)LOGERR(klogErr, rc, "Failed updating Alignment data in sequence table");
break;
}
KLoadProgressbar_Process(ctx->progress[ctx->pass - 1], 1, false);
}
MMArrayLock(ctx->id2value);
return rc;
}
static rc_t AlignmentUpdateSpotInfo(context_t *ctx, Alignment *align)
{
rc_t rc;
uint64_t keyId;
++ctx->pass;
KLoadProgressbar_Append(ctx->progress[ctx->pass - 1], ctx->alignCount);
rc = AlignmentStartUpdatingSpotIds(align);
while (rc == 0 && (rc = Quitting()) == 0) {
ctx_value_t *value;
rc = AlignmentGetSpotKey(align, &keyId);
if (rc) {
if (GetRCObject(rc) == rcRow && GetRCState(rc) == rcNotFound)
rc = 0;
break;
}
//assert(keyId >> 32 < ctx->keyToID.key2id_count);
//assert((uint32_t)keyId < ctx->keyToID.idCount[keyId >> 32]);
rc = MMArrayGet(ctx->id2value, (void **)&value, keyId);
if (rc == 0) {
int64_t const spotId = CTX_VALUE_GET_S_ID(*value);
if (spotId == 0) {
rc = RC(rcApp, rcTable, rcWriting, rcConstraint, rcViolated);
(void)PLOGERR(klogErr, (klogErr, rc, "Spot '$(id)' was never assigned a spot id, probably has no primary alignments", "id=%lx", keyId));
break;
}
#ifndef NO_METADATA
uint32_t group_id = keyId >> GROUPID_SHIFT;
auto [metadata, row_id] = ctx->m_read_groups[group_id]->metadata_by_key(keyId & KEYID_MASK);
auto spot_id = metadata->get<u64_t>(metadata_t::e_spotId).get(row_id);
if (spot_id != spotId) {
spdlog::error("Conflict: {} != {}", spot_id, spotId);
}
#endif
rc = AlignmentWriteSpotId(align, spotId);
}
KLoadProgressbar_Process(ctx->progress[ctx->pass - 1], 1, false);
}
MMArrayLock(ctx->id2value);
return rc;
}
#else
static rc_t WriteSoloFragments(context_t *ctx, Sequence *seq)
{
spdlog::stopwatch sw;
rc_t rc;
KDataBuffer fragBuf;
SequenceRecordStorage srecStorage;
SequenceRecord srec;
++ctx->pass;
memset(&srec, 0, sizeof(srec));
srec.ti = srecStorage.ti;
srec.readStart = srecStorage.readStart;
srec.readLen = srecStorage.readLen;
srec.orientation = srecStorage.orientation;
srec.is_bad = srecStorage.is_bad;
srec.alignmentCount = srecStorage.alignmentCount;
srec.aligned = srecStorage.aligned;
srec.cskey = srecStorage. cskey;
rc = KDataBufferMake(&fragBuf, 8, 0);
if (rc) {
(void)LOGERR(klogErr, rc, "KDataBufferMake failed");
return rc;
}
uint64_t idCount = 0;
for(const auto& rg : ctx->m_read_groups)
idCount += rg->m_total_spots;
KLoadProgressbar_Append(ctx->progress[ctx->pass - 1], idCount);
unsigned group_id = 0;
for (auto& gr : ctx->m_read_groups) {
gr->visit_metadata([&](metadata_t& metadata, unsigned group_id, size_t offset) {
size_t row_id = 0;
auto& fragCol = metadata.get<u32_t>(metadata_t::e_fragmentId);
auto fragment_it = fragCol.begin();
while (fragment_it.valid()) {
KLoadProgressbar_Process(ctx->progress[ctx->pass - 1], 1, false);
uint64_t keyId = ((uint64_t)group_id << GROUPID_SHIFT) | (offset + row_id);
#ifdef HAS_CTX_VALUE
ctx_value_t *value;
MMArrayGet(ctx->id2value, (void **)&value, keyId);
if (fragment_it.value() != value->fragmentId) {
spdlog::error("Solo fragment mismatch: {} != {}", fragment_it.value(), value->fragmentId);
}
#endif
if (fragment_it.value() != 0) {
size_t rsize;
size_t sz;
char const *src;
FragmentInfo const *fip;
rc = MemBankSize(ctx->frags, fragment_it.value(), &sz);
if (rc) {
(void)LOGERR(klogErr, rc, "KMemBankSize failed");
break;
}
rc = KDataBufferResize(&fragBuf, (size_t)sz);
if (rc) {
(void)LOGERR(klogErr, rc, "KDataBufferResize failed");
break;
}
rc = MemBankRead(ctx->frags, fragment_it.value(), 0, fragBuf.base, sz, &rsize);
if (rc) {
(void)LOGERR(klogErr, rc, "KMemBankRead failed");
break;
}
assert( rsize == sz );
fip = (const FragmentInfo*)fragBuf.base;
src = (char const *)&fip[1];
memset(&srecStorage, 0, sizeof(srecStorage));
if (metadata.get<bit_t>(metadata_t::e_unmated).test(row_id)) { //value->unmated
srec.numreads = 1;
srec.readLen[0] = fip->readlen;
srec.ti[0] = fip->ti;
srec.aligned[0] = fip->aligned;
srec.is_bad[0] = fip->is_bad;
srec.orientation[0] = fip->orientation;
srec.cskey[0] = fip->cskey;
} else {
unsigned const read = ((fip->aligned && metadata.get<u64_t>(metadata_t::E_PRIM_ID[0]).get_no_check(row_id) == 0) || metadata.get<bit_t>(metadata_t::e_unaligned_2).test(row_id))
? 1 : 0;
srec.numreads = 2;
srec.readLen[read] = fip->readlen;
srec.readStart[1] = srec.readLen[0];
srec.ti[read] = fip->ti;
srec.aligned[read] = fip->aligned;
srec.is_bad[read] = fip->is_bad;
srec.orientation[read] = fip->orientation;
srec.cskey[0] = srec.cskey[1] = 'N';
srec.cskey[read] = fip->cskey;
}
srec.seq = (char *)src;
srec.qual = (uint8_t *)(src + fip->readlen);
srec.spotGroup = (char *)(src + 2 * fip->readlen);
srec.spotGroupLen = fip->sglen;
srec.linkageGroup = (char *)(src + 2 * fip->readlen * fip->sglen);
srec.linkageGroupLen = fip->lglen;
srec.keyId = keyId;
INSDC_SRA_platform_id platform_id = ctx->m_isSingleGroup ?
metadata.get<u16_t>(metadata_t::e_platform).get(row_id) : ctx->m_read_groups[group_id]->m_platform;
rc = SequenceWriteRecord(seq, &srec, ctx->isColorSpace, metadata.get<bit_t>(metadata_t::e_pcr_dup).test(row_id), platform_id);
if (rc) {
(void)LOGERR(klogErr, rc, "SequenceWriteRecord failed");
break;
}
assert(metadata.get<u64_t>(metadata_t::e_spotId).get_no_check(row_id) == 0);
metadata.get<u64_t>(metadata_t::e_spotId).set(row_id, ++ctx->spotId);
#ifdef HAS_CTX_VALUE
CTX_VALUE_SET_S_ID(*value, ctx->spotId);
#endif
}
fragment_it.advance();
++row_id;
}
}, group_id);
++group_id;
}
/*
ctx->visit_metadata([&](metadata_t& metadata, unsigned group_id, size_t offset) {
size_t row_id = 0;
auto& fragCol = metadata.get<u32_t>(metadata_t::e_fragmentId);
auto fragment_it = fragCol.begin();
while (fragment_it.valid()) {
KLoadProgressbar_Process(ctx->progress[ctx->pass - 1], 1, false);
uint64_t keyId = ((uint64_t)group_id << GROUPID_SHIFT) | (offset + row_id);
#ifdef HAS_CTX_VALUE
ctx_value_t *value;
MMArrayGet(ctx->id2value, (void **)&value, keyId);
if (fragment_it.value() != value->fragmentId) {
spdlog::error("Solo fragment mismatch: {} != {}", fragment_it.value(), value->fragmentId);
}
#endif
if (fragment_it.value() != 0) {
size_t rsize;
size_t sz;
char const *src;
FragmentInfo const *fip;
rc = MemBankSize(ctx->frags, fragment_it.value(), &sz);
if (rc) {
(void)LOGERR(klogErr, rc, "KMemBankSize failed");
break;
}
rc = KDataBufferResize(&fragBuf, (size_t)sz);
if (rc) {
(void)LOGERR(klogErr, rc, "KDataBufferResize failed");
break;
}
rc = MemBankRead(ctx->frags, fragment_it.value(), 0, fragBuf.base, sz, &rsize);
if (rc) {
(void)LOGERR(klogErr, rc, "KMemBankRead failed");
break;
}
assert( rsize == sz );
fip = (const FragmentInfo*)fragBuf.base;
src = (char const *)&fip[1];
memset(&srecStorage, 0, sizeof(srecStorage));
if (metadata.get<bit_t>(metadata_t::e_unmated).test(row_id)) { //value->unmated
srec.numreads = 1;
srec.readLen[0] = fip->readlen;
srec.ti[0] = fip->ti;
srec.aligned[0] = fip->aligned;
srec.is_bad[0] = fip->is_bad;
srec.orientation[0] = fip->orientation;
srec.cskey[0] = fip->cskey;
} else {
unsigned const read = ((fip->aligned && metadata.get<u64_t>(metadata_t::E_PRIM_ID[0]).get_no_check(row_id) == 0) || metadata.get<bit_t>(metadata_t::e_unaligned_2).test(row_id))
? 1 : 0;
srec.numreads = 2;
srec.readLen[read] = fip->readlen;
srec.readStart[1] = srec.readLen[0];
srec.ti[read] = fip->ti;
srec.aligned[read] = fip->aligned;
srec.is_bad[read] = fip->is_bad;
srec.orientation[read] = fip->orientation;
srec.cskey[0] = srec.cskey[1] = 'N';
srec.cskey[read] = fip->cskey;
}
srec.seq = (char *)src;
srec.qual = (uint8_t *)(src + fip->readlen);
srec.spotGroup = (char *)(src + 2 * fip->readlen);
srec.spotGroupLen = fip->sglen;
srec.linkageGroup = (char *)(src + 2 * fip->readlen * fip->sglen);
srec.linkageGroupLen = fip->lglen;
srec.keyId = keyId;
INSDC_SRA_platform_id platform_id = ctx->m_isSingleGroup ?
metadata.get<u16_t>(metadata_t::e_platform).get(row_id) : ctx->m_read_groups[group_id]->m_platform;
rc = SequenceWriteRecord(seq, &srec, ctx->isColorSpace, metadata.get<bit_t>(metadata_t::e_pcr_dup).test(row_id), platform_id);
if (rc) {
(void)LOGERR(klogErr, rc, "SequenceWriteRecord failed");
break;
}
assert(metadata.get<u64_t>(metadata_t::e_spotId).get_no_check(row_id) == 0);
metadata.get<u64_t>(metadata_t::e_spotId).set(row_id, ++ctx->spotId);
#ifdef HAS_CTX_VALUE
CTX_VALUE_SET_S_ID(*value, ctx->spotId);
#endif
}
fragment_it.advance();
++row_id;
}
});
*/
KDataBufferWhack(&fragBuf);
ctx->clear_column<u32_t>(metadata_t::e_fragmentId);
ctx->clear_column<u16_t>(metadata_t::e_fragment_len1);
ctx->clear_column<u16_t>(metadata_t::e_fragment_len2);
ctx->clear_column<u16_t>(metadata_t::e_platform);
ctx->clear_column<bit_t>(metadata_t::e_pcr_dup);
spdlog::info("Solo fragments: {:.3} sec, memory: {:L}", sw, getCurrentRSS());
return rc;
}
static rc_t SequenceUpdateAlignInfo(context_t *ctx, Sequence *seq)
{
spdlog::stopwatch sw;
rc_t rc = 0;
uint64_t row;
uint64_t keyId;
++ctx->pass;
if (G.mode != mode_Remap) {
spdlog::info("Extraction start, memory: {:L}", getCurrentRSS());
ctx->extract_spotid();
spdlog::info("Extraction stop: {:.3} sec, memory: {:L}", sw, getCurrentRSS());
sw.reset();
}
size_t row_offset = 1;
KLoadProgressbar_Append(ctx->progress[ctx->pass - 1], ctx->spotId + 1);
typedef struct {
vector<uint64_t> keys;
vector<uint8_t> alignmentCount;//(BUFFER_SIZE * 2);
vector<int64_t> primaryId;//(BUFFER_SIZE * 2);
vector<uint8_t> unmated;//(BUFFER_SIZE * 2);
size_t offset = 0;
} key_batch_t;
atomic<bool> queue_done{false};
atomic<bool> exit_on_error{false};
atomic<bool> gather_done{false};
atomic<size_t> num_gathered = 0;
atomic<size_t> num_updated = 0;
size_t batches_processed = 0;
size_t batches_gathered = 0;
size_t batches_updated = 0;
ReaderWriterQueue<key_batch_t> gather_queue{12};
ReaderWriterQueue<key_batch_t> update_queue{4};
constexpr int BUFFER_SIZE = 10e6;
mutex metadata_mutex; // protects metadata in Remap mode
int const logLevel = klogWarn; /*G.assembleWithSecondary ? klogWarn : klogErr;*/
/* Two tasks and two queues
* Main thread puts keyIds into gather_queue
* Gather task works on gather_queue, extracts metadata and pus them into update_queue
* Update task works on update queue and updates VDB
*
*/
size_t expected_batches = (ctx->spotId/BUFFER_SIZE) + 1;
auto gather_task = ctx->m_executor->async([&]() {
key_batch_t batch;
while (exit_on_error == false) {
if (gather_queue.try_dequeue(batch)) {
++batches_gathered;
spdlog::stopwatch sw;
auto sz = batch.keys.size();
int page_size = sz/ctx->m_executor->num_workers();
int num_pages = page_size ? sz/page_size + 1 : 1;
if (num_pages == 1)
page_size = sz;
batch.alignmentCount.resize(sz * 2);
batch.primaryId.resize(sz * 2);
batch.unmated.resize(sz);
tf::Taskflow taskflow;
taskflow.for_each_index(0, num_pages, 1, [&](int index) {
size_t row_b = index * page_size;
size_t row_e = min<size_t>(row_b + page_size, batch.keys.size());
for (; row_b < row_e; ++row_b) {
++num_gathered;
auto keyId = batch.keys[row_b];
uint32_t group_id = keyId >> GROUPID_SHIFT;
uint64_t row_id = keyId & KEYID_MASK;
auto [metadata, local_row_id] = ctx->m_read_groups[group_id]->metadata_by_key(row_id);
batch.primaryId[row_b * 2] = (int64_t)metadata->get<u64_t>(metadata_t::E_PRIM_ID[0]).get_no_check(local_row_id);
batch.primaryId[row_b * 2 + 1] = (int64_t)metadata->get<u64_t>(metadata_t::E_PRIM_ID[1]).get_no_check(local_row_id);
batch.alignmentCount[row_b * 2] = (uint8_t)metadata->get<u16_t>(metadata_t::E_ALN_COUNT[0]).get_no_check(local_row_id);
batch.alignmentCount[row_b * 2 + 1] = (uint8_t)metadata->get<u16_t>(metadata_t::E_ALN_COUNT[1]).get_no_check(local_row_id);
batch.unmated[row_b] = metadata->get<bit_t>(metadata_t::e_unmated).test(local_row_id) ? 1 : 0;
if (G.mode == mode_Remap) {
const lock_guard<mutex> lock(metadata_mutex);
metadata->get<u64_t>(metadata_t::e_spotId).set(local_row_id, row_b + row_offset);
}
}
});
ctx->m_executor->run(taskflow).wait();
while (!update_queue.try_enqueue(move(batch))) {
if (exit_on_error)
break;
};
} else if (batches_gathered >= expected_batches) {
break;
}
}
gather_done = true;
});
auto update_task = ctx->m_executor->async([&]() {
key_batch_t batch;
rc = 0;
while (true) {
if (update_queue.try_dequeue(batch)) {
++batches_updated;
spdlog::stopwatch sw;
for (size_t i = 0; i < batch.keys.size(); ++i) {
++num_updated;
auto i_row = i + batch.offset;
if (batch.primaryId[i * 2] == 0 && batch.alignmentCount[i * 2] != 0) {
rc = RC(rcApp, rcTable, rcWriting, rcConstraint, rcViolated);
(void)PLOGERR(logLevel, (logLevel, rc, "Spot id $(id) read 1 never had a primary alignment", "id=%lx", batch.keys[i]));
}
bool is_unmated = batch.unmated[i];
if (!is_unmated && batch.primaryId[i * 2 + 1] == 0 && batch.alignmentCount[i * 2 + 1] != 0) {
rc = RC(rcApp, rcTable, rcWriting, rcConstraint, rcViolated);
(void)PLOGERR(logLevel, (logLevel, rc, "Spot id $(id) read 2 never had a primary alignment", "id=%lx", batch.keys[i]));
}
if (rc != 0 && logLevel == klogErr) {
exit_on_error = true;
break;
}
rc = SequenceUpdateAlignData(seq, i_row, is_unmated ? 1 : 2, &batch.primaryId[i * 2], &batch.alignmentCount[i * 2]);
if (rc) {
exit_on_error = true;
(void)LOGERR(klogErr, rc, "Failed updating Alignment data in sequence table");
break;
}
}
//spdlog::info("Finished updating batch {} in {:3} sec", batch.keys.size(), sw);
} else if (batches_updated >= expected_batches) {
break;
}
}
});
vector<uint64_t> keys;
keys.reserve(BUFFER_SIZE);
for (row = 1; row <= ctx->spotId; ++row) {
rc = SequenceReadKey(seq, row, &keyId);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "Failed to get key for row $(row)", "row=%u", (unsigned)row));
break;
}
if (G.mode != mode_Remap) {
auto spotId = ctx->m_spot_id_buffer[uint32_t(keyId >> GROUPID_SHIFT)].get(keyId & KEYID_MASK);
if (row != spotId) {
//if (row != metadata->get<u64_t>(metadata_t::e_spotId).get_no_check(local_row_id)) {
// auto spotId = metadata->get<u64_t>(metadata_t::e_spotId).get_no_check(local_row_id);
rc = RC(rcApp, rcTable, rcWriting, rcData, rcUnexpected);
(void)PLOGMSG(klogErr, (klogErr, "Unexpected spot id $(spotId) for row $(row), index $(idx)", "spotId=%u,row=%u,idx=%u", (unsigned)spotId, (unsigned)row, (unsigned)keyId));
break;
}
}
keys.push_back(keyId);
if (keys.size() == BUFFER_SIZE) {
key_batch_t batch;
batch.offset = row_offset;
row_offset += keys.size();
batch.keys = move(keys);
keys.clear();
keys.reserve(BUFFER_SIZE);
while (gather_queue.try_enqueue(move(batch)) == false) {
if (exit_on_error)
break;
};
++batches_processed;
if (exit_on_error)
break;
KLoadProgressbar_Process(ctx->progress[ctx->pass - 1], 1, false);
}
}
if (!keys.empty() && exit_on_error == false) {
key_batch_t batch;
batch.offset = row_offset;
row_offset += keys.size();
batch.keys = move(keys);
keys.clear();
while (gather_queue.try_enqueue(move(batch)) == false) {
if (exit_on_error)
break;
};
++batches_processed;
}
//while (gather_queue.peek() != nullptr)
// std::this_thread::sleep_for(std::chrono::milliseconds(100));
assert(gather_task.valid());
queue_done = true;
gather_task.get();
assert(update_task.valid());
update_task.get();
assert(exit_on_error || num_gathered == num_updated);
assert(exit_on_error || batches_processed == expected_batches);
assert(exit_on_error || batches_gathered == expected_batches);
assert(exit_on_error || batches_updated == expected_batches);
spdlog::info("Gathered: {:L}, updated: {:L}", num_gathered, num_updated);
spdlog::info("Queued: {:L}, Dequeued: {:L}", batches_gathered, batches_updated);
spdlog::info("Align Info: {:.3} sec, memory: {:L}", sw, getCurrentRSS());
return rc;
}
static rc_t AlignmentUpdateSpotInfo(context_t *ctx, Alignment *align)
{
spdlog::stopwatch sw;
rc_t rc;
uint64_t keyId;
++ctx->pass;
KLoadProgressbar_Append(ctx->progress[ctx->pass - 1], ctx->alignCount);
rc = AlignmentStartUpdatingSpotIds(align);
if (ctx->m_spot_id_buffer.empty())
ctx->extract_spotid();
while (rc == 0 && (rc = Quitting()) == 0) {
rc = AlignmentGetSpotKey(align, &keyId);
if (rc) {
if (GetRCObject(rc) == rcRow && GetRCState(rc) == rcNotFound)
rc = 0;
break;
}
uint32_t group_id = keyId >> GROUPID_SHIFT;
auto row_id = keyId & KEYID_MASK;
auto spotId = ctx->m_spot_id_buffer[group_id].get(row_id);
if (spotId == 0) {
rc = RC(rcApp, rcTable, rcWriting, rcConstraint, rcViolated);
(void)PLOGERR(klogErr, (klogErr, rc, "Spot '$(id)' was never assigned a spot id, probably has no primary alignments", "id=%lx", keyId));
break;
}
rc = AlignmentWriteSpotId(align, spotId);
KLoadProgressbar_Process(ctx->progress[ctx->pass - 1], 1, false);
}
spdlog::info("Align Update Spot Info: {:.3} sec", sw);
return rc;
}
#endif
static rc_t ArchiveBAM(VDBManager *mgr, VDatabase *db,
unsigned bamFiles, char const *bamFile[],
unsigned seqFiles, char const *seqFile[],
bool *has_alignments,
bool continuing)
{
std::locale::global(std::locale("C")); // enable comma as thousand separator
if (G.hasExtraLogging) {
auto logger = spdlog::stderr_logger_mt("stderr"); // send log to stderr
spdlog::set_default_logger(logger);
} else {
auto logger = spdlog::null_logger_mt("null"); // send log to stderr
spdlog::set_default_logger(logger);
}
spdlog::set_pattern("[%Y-%m-%d %H:%M:%S.%e] [%l] %v"); // default logging pattern (datetime, error level, error text)
spdlog::info("SIMD code = {}", bm::simd_version());
spdlog::info("Num threads = {}", G.numThreads);
spdlog::info("Search batch size = {}", G.searchBatchSize);
rc_t rc = 0;
rc_t rc2;
Reference ref;
Sequence seq;
Alignment *align;
static context_t *ctx = &GlobalContext;
bool has_sequences = false;
unsigned i;
*has_alignments = false;
rc = ReferenceInit(&ref, mgr, db);
if (rc)
return rc;
if (G.onlyVerifyReferences) {
for (i = 0; i < bamFiles && rc == 0; ++i) {
rc = ProcessBAM(bamFile[i], NULL, db, &ref, NULL, NULL, NULL, NULL);
}
ReferenceWhack(&ref, false);
return rc;
}
SequenceInit(&seq, db);
align = AlignmentMake(db);
ctx->m_inputSize = 0;
for (i = 0; i < bamFiles && rc == 0; ++i) {
if (strcmp(bamFile[i], "/dev/stdin") == 0) {
ctx->m_inputSize = 0;
break;
}
FILE *in = fopen(bamFile[i], "rb");
if (in) {
fseek(in, 0, SEEK_END);
ctx->m_inputSize += ftell(in);
fclose(in);
}
}
spdlog::info("Number of files: {}, Total size: {:L}", bamFiles, ctx->m_inputSize);
ctx->m_calcBatchSize = ctx->m_inputSize > 0;
rc = SetupContext(ctx, bamFiles + seqFiles);
if (rc)
return rc;
ctx->pass = 1;
for (i = 0; i < bamFiles && rc == 0; ++i) {
bool this_has_alignments = false;
bool this_has_sequences = false;
rc = ProcessBAM(bamFile[i], ctx, db, &ref, &seq, align, &this_has_alignments, &this_has_sequences);
*has_alignments |= this_has_alignments;
has_sequences |= this_has_sequences;
}
for (i = 0; i < seqFiles && rc == 0; ++i) {
bool this_has_alignments = false;
bool this_has_sequences = false;
rc = ProcessBAM(seqFile[i], ctx, db, &ref, &seq, align, &this_has_alignments, &this_has_sequences);
*has_alignments |= this_has_alignments;
has_sequences |= this_has_sequences;
}
spdlog::info("Processing done, memory: {:L}, spotCount: {:L}", getCurrentRSS(), ctx->spotId);
if (!continuing) {
ctx->release_search_memory();
// Clear the metadata columns that we don't need anymore
ctx->clear_column<bit_t>(metadata_t::e_unaligned_1);
ctx->clear_column<bit_t>(metadata_t::e_unaligned_2);
ctx->clear_column<bit_t>(metadata_t::e_hardclipped);
ctx->clear_column<bit_t>(metadata_t::e_primary_is_set);
spdlog::info("Spot assembly memory release, memory: {:L}", getCurrentRSS());
}
if (has_sequences) {
if (rc == 0 && (rc = Quitting()) == 0) {
if (G.mode == mode_Archive) {
(void)LOGMSG(klogInfo, "Writing unpaired sequences");
rc = WriteSoloFragments(ctx, &seq);
ContextReleaseMemBank(ctx);
}
if (rc == 0) {
rc = SequenceDoneWriting(&seq);
if (rc == 0) {
(void)LOGMSG(klogInfo, "Updating sequence alignment info");
rc = SequenceUpdateAlignInfo(ctx, &seq);
}
}
}
} else {
// Clear the metadata columns that we don't need anymore
ctx->clear_column<u64_t>(metadata_t::e_primaryId1);
ctx->clear_column<u64_t>(metadata_t::e_primaryId2);
ctx->clear_column<u32_t>(metadata_t::e_fragmentId);
ctx->clear_column<u16_t>(metadata_t::e_fragment_len1);
ctx->clear_column<u16_t>(metadata_t::e_fragment_len2);
ctx->clear_column<u16_t>(metadata_t::e_alignmentCount1);
ctx->clear_column<u16_t>(metadata_t::e_alignmentCount2);
ctx->clear_column<u16_t>(metadata_t::e_platform);
ctx->clear_column<bit_t>(metadata_t::e_pcr_dup);
}
if (*has_alignments && rc == 0 && (rc = Quitting()) == 0) {
(void)LOGMSG(klogInfo, "Writing alignment spot ids");
rc = AlignmentUpdateSpotInfo(ctx, align);
}
for (auto& b : ctx->m_spot_id_buffer) {
b.values.clear();
b.values.shrink_to_fit();
b.ext.clear();
b.ext.shrink_to_fit();
}
ctx->m_read_groups.clear();
ctx->m_executor.reset(nullptr);
spdlog::info("Whacking, memory: {:L}", getCurrentRSS());
rc2 = AlignmentWhack(align, *has_alignments && rc == 0 && (rc = Quitting()) == 0);
if (rc == 0)
rc = rc2;
rc2 = ReferenceWhack(&ref, *has_alignments && rc == 0 && (rc = Quitting()) == 0);
if (rc == 0)
rc = rc2;
SequenceWhack(&seq, rc == 0);
ContextRelease(ctx, continuing);
if (rc == 0) {
(void)LOGMSG(klogInfo, "Successfully loaded all files");
}
return rc;
}
rc_t WriteLoaderSignature(KMetadata *meta, char const progName[])
{
KMDataNode *node;
rc_t rc = KMetadataOpenNodeUpdate(meta, &node, "/");
if (rc == 0) {
rc = KLoaderMeta_Write(node, progName, __DATE__, "BAM", KAppVersion());
KMDataNodeRelease(node);
}
if (rc) {
(void)LOGERR(klogErr, rc, "Cannot update loader meta");
}
return rc;
}
rc_t OpenPath(char const path[], KDirectory **dir)
{
KDirectory *p;
rc_t rc = KDirectoryNativeDir(&p);
if (rc == 0) {
rc = KDirectoryOpenDirUpdate(p, dir, false, "%s", path);
KDirectoryRelease(p);
}
return rc;
}
static
rc_t ConvertDatabaseToUnmapped(VDatabase *db)
{
VTable* tbl;
rc_t rc = VDatabaseOpenTableUpdate(db, &tbl, "SEQUENCE");
if (rc == 0)
{
VTableRenameColumn(tbl, false, "CMP_ALTREAD", "ALTREAD");
VTableRenameColumn(tbl, false, "CMP_READ", "READ");
VTableRenameColumn(tbl, false, "CMP_ALTCSREAD", "ALTCSREAD");
VTableRenameColumn(tbl, false, "CMP_CSREAD", "CSREAD");
rc = VTableRelease(tbl);
}
return rc;
}
rc_t run(char const progName[],
unsigned bamFiles, char const *bamFile[],
unsigned seqFiles, char const *seqFile[],
bool continuing)
{
VDBManager *mgr;
rc_t rc;
rc_t rc2;
char const *db_type = G.expectUnsorted ? "NCBI:align:db:alignment_unsorted" : "NCBI:align:db:alignment_sorted";
rc = VDBManagerMakeUpdate(&mgr, NULL);
if (rc) {
(void)LOGERR (klogErr, rc, "failed to create VDB Manager!");
}
else {
bool has_alignments = false;
/* VDBManagerDisableFlushThread(mgr); */
rc = VDBManagerDisablePagemapThread(mgr);
if (rc == 0)
{
if (G.onlyVerifyReferences) {
rc = ArchiveBAM(mgr, NULL, bamFiles, bamFile, 0, NULL, &has_alignments, continuing);
}
else {
VSchema *schema;
rc = VDBManagerMakeSchema(mgr, &schema);
if (rc) {
(void)LOGERR (klogErr, rc, "failed to create schema");
}
else {
(void)(rc = VSchemaAddIncludePath(schema, "%s", G.schemaIncludePath));
rc = VSchemaParseFile(schema, "%s", G.schemaPath);
if (rc) {
(void)PLOGERR(klogErr, (klogErr, rc, "failed to parse schema file $(file)", "file=%s", G.schemaPath));
}
else {
VDatabase *db;
rc = VDBManagerCreateDB(mgr, &db, schema, db_type,
kcmInit + kcmMD5, "%s", G.outpath);
VSchemaRelease(schema);
if (rc == 0) {
rc = ArchiveBAM(mgr, db, bamFiles, bamFile, seqFiles, seqFile, &has_alignments, continuing);
if (rc == 0)
PrintChangeReport();
if (rc == 0 && !has_alignments) {
rc = ConvertDatabaseToUnmapped(db);
}
else if (rc == 0 && lmc != NULL) {
VTable *tbl = NULL;
KTable *ktbl = NULL;
KMetadata *meta = NULL;
KMDataNode *node = NULL;
VDatabaseOpenTableUpdate(db, &tbl, "REFERENCE");
VTableOpenKTableUpdate(tbl, &ktbl);
VTableRelease(tbl);
KTableOpenMetadataUpdate(ktbl, &meta);
KTableRelease(ktbl);
KMetadataOpenNodeUpdate(meta, &node, "LOW_MATCH_COUNT");
KMetadataRelease(meta);
RecordLowMatchCounts(node);
KMDataNodeRelease(node);
LowMatchCounterFree(lmc);
lmc = NULL;
}
VDatabaseRelease(db);
if (rc == 0 && G.globalMode == mode_Remap && !continuing) {
VTable *tbl = NULL;
VDBManagerOpenDBUpdate(mgr, &db, NULL, G.firstOut);
VDatabaseOpenTableUpdate(db, &tbl, "SEQUENCE");
VDatabaseRelease(db);
VTableDropColumn(tbl, "TMP_KEY_ID");
VTableDropColumn(tbl, "READ");
VTableDropColumn(tbl, "ALTREAD");
VTableRelease(tbl);
}
if (rc == 0) {
KMetadata *meta = NULL;
{
KDBManager *kmgr = NULL;
rc = VDBManagerOpenKDBManagerUpdate(mgr, &kmgr);
if (rc == 0) {
KDatabase *kdb;
rc = KDBManagerOpenDBUpdate(kmgr, &kdb, "%s", G.outpath);
if (rc == 0) {
rc = KDatabaseOpenMetadataUpdate(kdb, &meta);
KDatabaseRelease(kdb);
}
KDBManagerRelease(kmgr);
}
}
if (rc == 0) {
rc = WriteLoaderSignature(meta, progName);
if (rc == 0) {
KMDataNode *changes = NULL;
rc = KMetadataOpenNodeUpdate(meta, &changes, "CHANGES");
if (rc == 0)
RecordChanges(changes, "CHANGE");
KMDataNodeRelease(changes);
}
KMetadataRelease(meta);
}
}
}
}
}
}
}
rc2 = VDBManagerRelease(mgr);
if (rc2)
(void)LOGERR(klogWarn, rc2, "Failed to release VDB Manager");
if (rc == 0)
rc = rc2;
}
return rc;
}
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