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#include <algorithm>
#include <cctype>
#include <cstdio>
#include <fstream>
#include <iostream>
#include <iterator>
#include <memory>
#include <mutex>
#include <random>
#include <type_traits>
#include <unordered_map>
#include <vector>
#include "tclap/CmdLine.h"
#include <cereal/archives/binary.hpp>
#include <cereal/archives/json.hpp>
#include <cereal/types/string.hpp>
#include <cereal/types/unordered_map.hpp>
#include <cereal/types/utility.hpp>
#include <cereal/types/vector.hpp>
#include "BooMap.hpp"
#include "xxhash.h"
#include "spdlog/spdlog.h"
#include "FastxParser.hpp"
// Jellyfish 2 include
#include "jellyfish/mer_dna.hpp"
#include "divsufsort.h"
#include "divsufsort64.h"
#include "RapMapFileSystem.hpp"
#include "RapMapUtils.hpp"
#include "ScopedTimer.hpp"
#include "bit_array.h"
#include "JFRaw.hpp"
#include "jellyfish/binary_dumper.hpp"
#include "jellyfish/file_header.hpp"
#include "jellyfish/hash_counter.hpp"
#include "jellyfish/mer_iterator.hpp"
#include "jellyfish/mer_overlap_sequence_parser.hpp"
#include "jellyfish/thread_exec.hpp"
#include "rank9b.h"
#include "sparsehash/dense_hash_map"
#include "IndexHeader.hpp"
#include <chrono>
using single_parser = fastx_parser::FastxParser<fastx_parser::ReadSeq>;
using TranscriptID = uint32_t;
using TranscriptIDVector = std::vector<TranscriptID>;
using KmerIDMap = std::vector<TranscriptIDVector>;
using MerMapT = jellyfish::cooperative::hash_counter<rapmap::utils::my_mer>;
bool buildSA(const std::string& outputDir, std::string& concatText, size_t tlen,
std::vector<int64_t>& SA) {
// IndexT is the signed index type
// UIndexT is the unsigned index type
using IndexT = int64_t;
using UIndexT = uint64_t;
bool success{false};
std::ofstream saStream(outputDir + "sa.bin", std::ios::binary);
{
ScopedTimer timer;
SA.resize(tlen, 0);
IndexT textLen = static_cast<IndexT>(tlen);
std::cerr << "Building suffix array . . . ";
auto ret = divsufsort64(
reinterpret_cast<unsigned char*>(const_cast<char*>(concatText.data())),
SA.data(), tlen);
success = (ret == 0);
if (success) {
std::cerr << "success\n";
{
ScopedTimer timer2;
std::cerr << "saving to disk . . . ";
cereal::BinaryOutputArchive saArchive(saStream);
saArchive(SA);
std::cerr << "done\n";
}
} else {
std::cerr << "FAILURE: return code from libdivsufsort64() was " << ret
<< "\n";
saStream.close();
std::exit(1);
}
std::cerr << "done\n";
}
saStream.close();
return success;
}
// IndexT is the index type.
// int32_t for "small" suffix arrays
// int64_t for "large" ones
template <typename IndexT>
bool buildPerfectHash(const std::string& outputDir, std::string& concatText,
size_t tlen, uint32_t k, std::vector<IndexT>& SA,
uint32_t numHashThreads) {
BooMap<uint64_t, rapmap::utils::SAInterval<IndexT>> intervals;
// The start and stop of the current interval
IndexT start = 0, stop = 0;
// An iterator to the beginning of the text
auto textB = concatText.begin();
auto textE = concatText.end();
// The current k-mer as a string
rapmap::utils::my_mer mer;
bool currentValid{false};
std::string currentKmer;
std::string nextKmer;
while (stop < tlen) {
// Check if the string starting at the
// current position is valid (i.e. doesn't contain $)
// and is <= k bases from the end of the string
nextKmer = concatText.substr(SA[stop], k);
if (nextKmer.length() == k and
nextKmer.find_first_of('$') == std::string::npos) {
// If this is a new k-mer, then hash the current k-mer
if (nextKmer != currentKmer) {
if (currentKmer.length() == k and
currentKmer.find_first_of('$') == std::string::npos) {
mer = rapmap::utils::my_mer(currentKmer);
auto bits = mer.get_bits(0, 2 * k);
intervals.add(std::move(bits), {start, stop});
// push_back(std::make_pair<uint64_t,
// rapmap::utils::SAInterval<IndexT>>(std::move(bits), {start,
// stop}));
}
currentKmer = nextKmer;
start = stop;
}
} else {
// If this isn't a valid suffix (contains a $)
// If the previous interval was valid, put it
// in the hash.
if (currentKmer.length() == k and
currentKmer.find_first_of('$') == std::string::npos) {
mer = rapmap::utils::my_mer(currentKmer);
auto bits = mer.get_bits(0, 2 * k);
// intervals.push_back(std::make_pair<uint64_t,
// rapmap::utils::SAInterval<IndexT>>(std::move(bits), {start, stop}));
intervals.add(std::move(bits), {start, stop});
}
// The current interval is invalid and empty
currentKmer = nextKmer;
start = stop;
}
if (stop % 1000000 == 0) {
std::cerr << "\r\rprocessed " << stop << " positions";
}
// We always update the end position
++stop;
}
if (start < tlen) {
if (currentKmer.length() == k and
currentKmer.find_first_of('$') != std::string::npos) {
mer = rapmap::utils::my_mer(currentKmer);
auto bits = mer.get_bits(0, 2 * k);
// intervals.push_back(std::make_pair<uint64_t,
// rapmap::utils::SAInterval<IndexT>>(std::move(bits), {start, stop}));
intervals.add(std::move(bits), {start, stop});
}
}
// std::cerr << "\nthere are " << intervals.size() << " intervals of the
// selected depth\n";
std::cout << "building perfect hash function\n";
intervals.build(numHashThreads);
std::cout << "\ndone.\n";
std::string outputPrefix = outputDir + "hash_info";
std::cout << "saving the perfect hash and SA intervals to disk ... ";
intervals.save(outputPrefix);
std::cout << "done.\n";
return true;
}
bool buildSA(const std::string& outputDir, std::string& concatText, size_t tlen,
std::vector<int32_t>& SA) {
// IndexT is the signed index type
// UIndexT is the unsigned index type
using IndexT = int32_t;
using UIndexT = uint32_t;
bool success{false};
std::ofstream saStream(outputDir + "sa.bin", std::ios::binary);
{
ScopedTimer timer;
SA.resize(tlen, 0);
IndexT textLen = static_cast<IndexT>(tlen);
std::cerr << "Building suffix array . . . ";
auto ret = divsufsort(
reinterpret_cast<unsigned char*>(const_cast<char*>(concatText.data())),
SA.data(), tlen);
success = (ret == 0);
if (success) {
std::cerr << "success\n";
{
ScopedTimer timer2;
std::cerr << "saving to disk . . . ";
cereal::BinaryOutputArchive saArchive(saStream);
saArchive(SA);
std::cerr << "done\n";
}
} else {
std::cerr << "FAILURE: return code from libdivsufsort() was " << ret
<< "\n";
saStream.close();
std::exit(1);
}
std::cerr << "done\n";
}
saStream.close();
return success;
}
// IndexT is the index type.
// int32_t for "small" suffix arrays
// int64_t for "large" ones
template <typename IndexT>
bool buildHash(const std::string& outputDir, std::string& concatText,
size_t tlen, uint32_t k, std::vector<IndexT>& SA) {
// Now, build the k-mer lookup table
google::dense_hash_map<uint64_t, rapmap::utils::SAInterval<IndexT>,
rapmap::utils::KmerKeyHasher>
khash;
khash.set_empty_key(std::numeric_limits<uint64_t>::max());
// The start and stop of the current interval
IndexT start = 0, stop = 0;
// An iterator to the beginning of the text
auto textB = concatText.begin();
auto textE = concatText.end();
// The current k-mer as a string
rapmap::utils::my_mer mer;
bool currentValid{false};
std::string currentKmer;
std::string nextKmer;
while (stop < tlen) {
// Check if the string starting at the
// current position is valid (i.e. doesn't contain $)
// and is <= k bases from the end of the string
nextKmer = concatText.substr(SA[stop], k);
if (nextKmer.length() == k and
nextKmer.find_first_of('$') == std::string::npos) {
// If this is a new k-mer, then hash the current k-mer
if (nextKmer != currentKmer) {
if (currentKmer.length() == k and
currentKmer.find_first_of('$') == std::string::npos) {
mer = rapmap::utils::my_mer(currentKmer);
auto bits = mer.get_bits(0, 2 * k);
auto hashIt = khash.find(bits);
if (hashIt == khash.end()) {
if (start > 1) {
if (concatText.substr(SA[start - 1], k) ==
concatText.substr(SA[start], k)) {
std::cerr << "T[SA[" << start - 1 << "]:" << k
<< "] = " << concatText.substr(SA[start - 1], k)
<< " = T[SA[" << start << "]:" << k << "]\n";
std::cerr << "start = " << start << ", stop = " << stop << "\n";
std::cerr << "[fatal (1)] THIS SHOULD NOT HAPPEN\n";
std::exit(1);
}
}
if (start == stop) {
std::cerr << "[fatal (2)] Interval is empty! (start = " << start
<< ") = (stop = " << stop << ")\n";
}
if (start == stop) {
std::cerr << "[fatal (3)] Interval is empty! (start = " << start
<< ") = (stop = " << stop << ")\n";
}
khash[bits] = {start, stop};
} else {
std::cerr << "\nERROR (1): trying to add same suffix "
<< currentKmer << " (len = " << currentKmer.length()
<< ") multiple times!\n";
auto prevInt = hashIt->second;
std::cerr << "existing interval is [" << prevInt.begin << ", "
<< prevInt.end << ")\n";
for (auto x = prevInt.begin; x < prevInt.end; ++x) {
auto suff = concatText.substr(SA[x], k);
for (auto c : suff) {
std::cerr << "*" << c << "*";
}
std::cerr << " (len = " << suff.length() << ")\n";
}
std::cerr << "new interval is [" << start << ", " << stop << ")\n";
for (auto x = start; x < stop; ++x) {
auto suff = concatText.substr(SA[x], k);
for (auto c : suff) {
std::cerr << "*" << c << "*";
}
std::cerr << "\n";
}
}
}
currentKmer = nextKmer;
start = stop;
}
} else {
// If this isn't a valid suffix (contains a $)
// If the previous interval was valid, put it
// in the hash.
if (currentKmer.length() == k and
currentKmer.find_first_of('$') == std::string::npos) {
mer = rapmap::utils::my_mer(currentKmer);
auto bits = mer.get_bits(0, 2 * k);
auto hashIt = khash.find(bits);
if (hashIt == khash.end()) {
if (start > 2) {
if (concatText.substr(SA[start - 1], k) ==
concatText.substr(SA[start], k)) {
std::cerr << "T[SA[" << start - 1 << "]:" << k
<< "] = " << concatText.substr(SA[start - 1], k)
<< " = T[SA[" << start << "]:" << k << "]\n";
std::cerr << "start = " << start << ", stop = " << stop << "\n";
std::cerr << "[fatal (4)] THIS SHOULD NOT HAPPEN\n";
std::exit(1);
}
}
khash[bits] = {start, stop};
} else {
std::cerr << "\nERROR (2): trying to add same suffix " << currentKmer
<< "multiple times!\n";
auto prevInt = hashIt->second;
std::cerr << "existing interval is [" << prevInt.begin << ", "
<< prevInt.end << ")\n";
for (auto x = prevInt.begin; x < prevInt.end; ++x) {
std::cerr << concatText.substr(SA[x], k) << "\n";
}
std::cerr << "new interval is [" << start << ", " << stop << ")\n";
for (auto x = start; x < stop; ++x) {
std::cerr << concatText.substr(SA[x], k) << "\n";
}
}
}
// The current interval is invalid and empty
currentKmer = nextKmer;
start = stop;
}
if (stop % 1000000 == 0) {
std::cerr << "\r\rprocessed " << stop << " positions";
}
// We always update the end position
++stop;
}
if (start < tlen) {
if (currentKmer.length() == k and
currentKmer.find_first_of('$') != std::string::npos) {
mer = rapmap::utils::my_mer(currentKmer);
khash[mer.get_bits(0, 2 * k)] = {start, stop};
}
}
std::cerr << "\nkhash had " << khash.size() << " keys\n";
std::ofstream hashStream(outputDir + "hash.bin", std::ios::binary);
{
ScopedTimer timer;
std::cerr << "saving hash to disk . . . ";
cereal::BinaryOutputArchive hashArchive(hashStream);
// hashArchive(k);
khash.serialize(typename google::dense_hash_map<
uint64_t, rapmap::utils::SAInterval<IndexT>,
rapmap::utils::KmerKeyHasher>::NopointerSerializer(),
&hashStream);
// hashArchive(khash);
std::cerr << "done\n";
}
hashStream.close();
return true;
}
// To use the parser in the following, we get "jobs" until none is
// available. A job behaves like a pointer to the type
// jellyfish::sequence_list (see whole_sequence_parser.hpp).
template <typename ParserT> //, typename CoverageCalculator>
void indexTranscriptsSA(ParserT* parser, std::string& outputDir,
bool noClipPolyA, bool usePerfectHash,
uint32_t numHashThreads,
std::string& sepStr,
std::mutex& iomutex,
std::shared_ptr<spdlog::logger> log) {
// Seed with a real random value, if available
std::random_device rd;
// Create a random uniform distribution
std::default_random_engine eng(rd());
std::uniform_int_distribution<> dis(0, 3);
uint32_t n{0};
uint32_t k = rapmap::utils::my_mer::k();
std::vector<std::string> transcriptNames;
std::vector<int64_t> transcriptStarts;
// std::vector<uint32_t> positionIDs;
constexpr char bases[] = {'A', 'C', 'G', 'T'};
uint32_t polyAClipLength{10};
uint32_t numPolyAsClipped{0};
uint32_t numNucleotidesReplaced{0};
std::string polyA(polyAClipLength, 'A');
using TranscriptList = std::vector<uint32_t>;
using eager_iterator = MerMapT::array::eager_iterator;
using KmerBinT = uint64_t;
bool clipPolyA = !noClipPolyA;
// http://biology.stackexchange.com/questions/21329/whats-the-longest-transcript-known
// longest human transcript is Titin (108861), so this gives us a *lot* of
// leeway before
// we issue any warning.
size_t tooLong = 200000;
size_t numDistinctKmers{0};
size_t numKmers{0};
size_t currIndex{0};
std::cerr << "\n[Step 1 of 4] : counting k-mers\n";
// rsdic::RSDicBuilder rsdb;
std::vector<uint64_t>
onePos; // Positions in the bit array where we should write a '1'
fmt::MemoryWriter txpSeqStream;
{
ScopedTimer timer;
// Get the read group by which this thread will
// communicate with the parser (*once per-thread*)
auto rg = parser->getReadGroup();
while (parser->refill(rg)) {
for (auto& read : rg) { // for each sequence
std::string& readStr = read.seq;
readStr.erase(
std::remove_if(readStr.begin(), readStr.end(),
[](const char a) -> bool { return !(isprint(a)); }),
readStr.end());
uint32_t readLen = readStr.size();
// First, replace non ATCG nucleotides
for (size_t b = 0; b < readLen; ++b) {
readStr[b] = ::toupper(readStr[b]);
int c = jellyfish::mer_dna::code(readStr[b]);
// Replace non-ACGT bases with pseudo-random bases
if (jellyfish::mer_dna::not_dna(c)) {
char rbase = bases[dis(eng)];
c = jellyfish::mer_dna::code(rbase);
readStr[b] = rbase;
++numNucleotidesReplaced;
}
}
// Now, do Kallisto-esque clipping of polyA tails
if (clipPolyA) {
if (readStr.size() > polyAClipLength and
readStr.substr(readStr.length() - polyAClipLength) == polyA) {
auto newEndPos = readStr.find_last_not_of("Aa");
// If it was all As
if (newEndPos == std::string::npos) {
log->warn("Entry with header [{}] appeared to be all A's; it "
"will be removed from the index!",
read.name);
readStr.resize(0);
} else {
readStr.resize(newEndPos + 1);
}
++numPolyAsClipped;
}
}
readLen = readStr.size();
// If the transcript was completely removed during clipping, don't
// include it in the index.
if (readStr.size() > 0) {
// If we're suspicious the user has fed in a *genome* rather
// than a transcriptome, say so here.
if (readStr.size() >= tooLong) {
log->warn("Entry with header [{}] was longer than {} nucleotides. "
"Are you certain that "
"we are indexing a transcriptome and not a genome?",
read.name, tooLong);
} else if (readStr.size() < k) {
log->warn("Entry with header [{}], had length less than "
"the k-mer length of {} (perhaps after poly-A clipping)",
read.name, k);
}
uint32_t txpIndex = n++;
// The name of the current transcript
auto& recHeader = read.name;
transcriptNames.emplace_back(
recHeader.substr(0, recHeader.find_first_of(sepStr)));//" \t")));
// The position at which this transcript starts
transcriptStarts.push_back(currIndex);
txpSeqStream << readStr;
txpSeqStream << '$';
currIndex += readLen + 1;
onePos.push_back(currIndex - 1);
} else {
log->warn("Discarding entry with header [{}], since it had length 0 "
"(perhaps after poly-A clipping)",
read.name);
}
}
if (n % 10000 == 0) {
std::cerr << "\r\rcounted k-mers for " << n << " transcripts";
}
}
}
std::cerr << "\n";
std::cerr << "Replaced " << numNucleotidesReplaced
<< " non-ATCG nucleotides\n";
std::cerr << "Clipped poly-A tails from " << numPolyAsClipped
<< " transcripts\n";
// Put the concatenated text in a string
std::string concatText = txpSeqStream.str();
// And clear the stream
txpSeqStream.clear();
// Build the suffix array
size_t tlen = concatText.length();
size_t maxInt = std::numeric_limits<int32_t>::max();
bool largeIndex = (tlen + 1 > maxInt);
// Make our dense bit arrray
BIT_ARRAY* bitArray = bit_array_create(concatText.length());
for (auto p : onePos) {
bit_array_set_bit(bitArray, p);
}
onePos.clear();
onePos.shrink_to_fit();
std::string rsFileName = outputDir + "rsd.bin";
FILE* rsFile = fopen(rsFileName.c_str(), "w");
{
ScopedTimer timer;
std::cerr << "Building rank-select dictionary and saving to disk ";
bit_array_save(bitArray, rsFile);
std::cerr << "done\n";
}
fclose(rsFile);
bit_array_free(bitArray);
std::ofstream seqStream(outputDir + "txpInfo.bin", std::ios::binary);
{
ScopedTimer timer;
std::cerr << "Writing sequence data to file . . . ";
cereal::BinaryOutputArchive seqArchive(seqStream);
seqArchive(transcriptNames);
if (largeIndex) {
seqArchive(transcriptStarts);
} else {
std::vector<int32_t> txpStarts(transcriptStarts.size(), 0);
size_t numTranscriptStarts = transcriptStarts.size();
for (size_t i = 0; i < numTranscriptStarts; ++i) {
txpStarts[i] = static_cast<int32_t>(transcriptStarts[i]);
}
transcriptStarts.clear();
transcriptStarts.shrink_to_fit();
{ seqArchive(txpStarts); }
}
// seqArchive(positionIDs);
seqArchive(concatText);
std::cerr << "done\n";
}
seqStream.close();
// clear stuff we no longer need
// positionIDs.clear();
// positionIDs.shrink_to_fit();
transcriptStarts.clear();
transcriptStarts.shrink_to_fit();
transcriptNames.clear();
transcriptNames.shrink_to_fit();
// done clearing
if (largeIndex) {
largeIndex = true;
std::cerr << "[info] Building 64-bit suffix array "
"(length of generalized text is "
<< tlen << " )\n";
using IndexT = int64_t;
std::vector<IndexT> SA;
bool success = buildSA(outputDir, concatText, tlen, SA);
if (!success) {
std::cerr << "[fatal] Could not build the suffix array!\n";
std::exit(1);
}
if (usePerfectHash) {
success = buildPerfectHash<IndexT>(outputDir, concatText, tlen, k, SA,
numHashThreads);
} else {
success = buildHash<IndexT>(outputDir, concatText, tlen, k, SA);
}
if (!success) {
std::cerr << "[fatal] Could not build the suffix interval hash!\n";
std::exit(1);
}
} else {
std::cerr << "[info] Building 32-bit suffix array "
"(length of generalized text is "
<< tlen << ")\n";
using IndexT = int32_t;
std::vector<IndexT> SA;
bool success = buildSA(outputDir, concatText, tlen, SA);
if (!success) {
std::cerr << "[fatal] Could not build the suffix array!\n";
std::exit(1);
}
if (usePerfectHash) {
success = buildPerfectHash<IndexT>(outputDir, concatText, tlen, k, SA,
numHashThreads);
} else {
success = buildHash<IndexT>(outputDir, concatText, tlen, k, SA);
}
if (!success) {
std::cerr << "[fatal] Could not build the suffix interval hash!\n";
std::exit(1);
}
}
std::string indexVersion = "q3";
IndexHeader header(IndexType::QUASI, indexVersion, true, k, largeIndex,
usePerfectHash);
// Finally (since everything presumably succeeded) write the header
std::ofstream headerStream(outputDir + "header.json");
{
cereal::JSONOutputArchive archive(headerStream);
archive(header);
}
headerStream.close();
}
int rapMapSAIndex(int argc, char* argv[]) {
std::cerr << "RapMap Indexer\n";
TCLAP::CmdLine cmd("RapMap Indexer");
TCLAP::ValueArg<std::string> transcripts("t", "transcripts",
"The transcript file to be indexed",
true, "", "path");
TCLAP::ValueArg<std::string> index(
"i", "index", "The location where the index should be written", true, "",
"path");
TCLAP::ValueArg<uint32_t> kval("k", "klen", "The length of k-mer to index",
false, 31, "positive integer less than 32");
TCLAP::ValueArg<std::string> customSeps("s", "headerSep", "Instead of a space or tab, break the header at the first "
"occurrence of this string, and name the transcript as the token before "
"the first separator", false, " \t", "string");
TCLAP::SwitchArg noClip(
"n", "noClip",
"Don't clip poly-A tails from the ends of target sequences", false);
TCLAP::SwitchArg perfectHash(
"p", "perfectHash", "Use a perfect hash instead of dense hash --- "
"somewhat slows construction, but uses less memory",
false);
TCLAP::ValueArg<uint32_t> numHashThreads(
"x", "numThreads",
"Use this many threads to build the perfect hash function", false, 4,
"positive integer <= # cores");
cmd.add(transcripts);
cmd.add(index);
cmd.add(kval);
cmd.add(noClip);
cmd.add(perfectHash);
cmd.add(customSeps);
cmd.add(numHashThreads);
cmd.parse(argc, argv);
// stupid parsing for now
std::string transcriptFile(transcripts.getValue());
std::vector<std::string> transcriptFiles({transcriptFile});
std::string sepStr = customSeps.getValue();
uint32_t k = kval.getValue();
if (k % 2 == 0) {
std::cerr << "Error: k must be an odd value, you chose " << k << '\n';
std::exit(1);
} else if (k > 31) {
std::cerr << "Error: k must not be larger than 31, you chose " << k << '\n';
std::exit(1);
}
rapmap::utils::my_mer::k(k);
std::string indexDir = index.getValue();
if (indexDir.back() != '/') {
indexDir += '/';
}
bool dirExists = rapmap::fs::DirExists(indexDir.c_str());
bool dirIsFile = rapmap::fs::FileExists(indexDir.c_str());
if (dirIsFile) {
std::cerr << "The requested index directory already exists as a file.";
std::exit(1);
}
if (!dirExists) {
rapmap::fs::MakeDir(indexDir.c_str());
}
std::string logPath = indexDir + "quasi_index.log";
auto fileSink = std::make_shared<spdlog::sinks::simple_file_sink_st>(logPath);
auto consoleSink = std::make_shared<spdlog::sinks::stderr_sink_st>();
auto consoleLog = spdlog::create("stderrLog", {consoleSink});
auto fileLog = spdlog::create("fileLog", {fileSink});
auto jointLog = spdlog::create("jointLog", {fileSink, consoleSink});
size_t numThreads{1};
std::unique_ptr<single_parser> transcriptParserPtr{nullptr};
size_t numProd = 1;
transcriptParserPtr.reset(
new single_parser(transcriptFiles, numThreads, numProd));
transcriptParserPtr->start();
bool noClipPolyA = noClip.getValue();
bool usePerfectHash = perfectHash.getValue();
uint32_t numPerfectHashThreads = numHashThreads.getValue();
std::mutex iomutex;
indexTranscriptsSA(transcriptParserPtr.get(), indexDir, noClipPolyA,
usePerfectHash, numPerfectHashThreads, sepStr, iomutex, jointLog);
return 0;
}
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