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/*
* Copyright (c) Medical Research Council 2002. All rights reserved.
*
* Permission to use, copy, modify and distribute this software and its
* documentation for any purpose is hereby granted without fee, provided that
* this copyright and notice appears in all copies.
*
* This file was written as part of the Staden Package at the MRC Laboratory
* of Molecular Biology, Hills Road, Cambridge, CB2 2QH, United Kingdom.
*
* MRC disclaims all warranties with regard to this software.
*
*/
#include <staden_config.h>
#include <cassert>
#include <new> // For bad_alloc
#include <cstdio> // For fprintf(), fflush(), etc
#include <cctype> // For isspace(), toupper()
#include <cstring> // For strcpy(), strcat(), etc
#include <cstdlib> // For atof()
#include <staden.h> // For io_lib and licence stuff
#include <mutlib.h> // For mutation library
#include <pathutil.h> // For MakeFullPath()
#include <stringlist.hpp> // For StringList
#include <string.h> // Needed for Solaris C++ compiler?
//-------------
// Module Data
//-------------
const int BUFSIZE = 512;
//------------------
// Helper Functions
//------------------
void PrintBanner()
{
std::fprintf( stdout, "Mutscan v1.00, by Mark Jordan.\n"
"MRC Laboratory of Molecular Biology, Copyright (C) 2002. All Rights Reserved.\n" );
std::fflush( stdout );
}
void PrintUsage( mutscan_t& ms )
{
std::fprintf( stdout, "Usage : mutscan [options] <experiment-files>\n"
"Options:\n"
"[-a<mutation-count>] = Alignment failure report threshold (default=%0.0f)\n"
"[-c] = Complement reverse strand tags (default=off)\n"
"[-f<file-of-filenames>] = File of experiment filenames\n"
"[-h<heterzygote-SNR>] = Heterzygote SNR threshold in dB (default=%0.2f)\n"
"[-l<peakdrop-threshold>] = Lower peak drop threshold percentage (default=%0.2f)\n"
"[-n<noise-threshold] = Noise level threshold percentage (default=%0.2f)\n"
"[-q] = Quiet mode (default=off)\n"
"[-u<peakdrop-threshold>] = Upper peak drop threshold percentage (default=%0.2f)\n"
"[-w<search-window-size>] = Peak search window size in bases (default=%0.2f)\n"
"[-p] = Proximity filter threshold... (default=7)\n"
"[-z] = Enter debugging loop and wait... (default=off)\n",
ms.Parameter[MUTSCAN_PARAMETER_ALIGNFAIL_THRESHOLD],
ms.Parameter[MUTSCAN_PARAMETER_HETSNR_THRESHOLD],
ms.Parameter[MUTSCAN_PARAMETER_PEAKDROP_LOWER],
ms.Parameter[MUTSCAN_PARAMETER_NOISE_THRESHOLD],
ms.Parameter[MUTSCAN_PARAMETER_PEAKDROP_UPPER],
ms.Parameter[MUTSCAN_PARAMETER_SEARCH_WINDOW]
);
std::fflush( stdout );
}
// Filter clusters of tags too close to the end of MCOV
void filter_tags ( mutscan_t& ms, int nTags, int threshold ) {
int *scores_a, *scores;
int cov_start = 0, cov_end = 0;
const int win_len = 5;
// Get coverage range
for( int i=0; i<nTags; i++ ) {
mutlib_tag_t* pTag = MutScanGetTag( &ms, i );
assert(pTag != NULL);
if (std::strcmp(pTag->Type,"MCOV") == 0) {
cov_start = pTag->Position[0];
cov_end = pTag->Position[1];
break;
}
}
printf("Cov=%d..%d\n", cov_start, cov_end);
/*
* Allocate an array indexed from -win_len to cov_end+win_len.
* Each tag gets a peak of width 2*win_len+1 centred on it and is
* accumulated in this array.
*/
scores_a = new int[win_len*2+cov_end+1];
scores = &scores_a[win_len];
memset(scores_a, 0, (win_len*2+cov_end) *sizeof(*scores_a));
// Convolve tag peaks
for (int i=0; i < nTags; i++) {
mutlib_tag_t* pTag = MutScanGetTag( &ms, i );
int pos = pTag->Position[0];
if (pos < 0) pos = 0;
if (pos > cov_end) pos = cov_end;
scores[pos] += win_len+1;
for (int j=1; j<=win_len; j++) {
scores[pos+j] += win_len+1 - j;
scores[pos-j] += win_len+1 - j;
}
}
// Filter tags
for (int i=0; i < nTags; i++) {
mutlib_tag_t* pTag = MutScanGetTag( &ms, i );
int pos = pTag->Position[0];
if (std::strcmp(pTag->Type,"MCOV") == 0)
continue;
if (scores[pos] >= threshold) {
pTag->Type[3] = '?';
}
}
delete [] scores_a;
#if 0
int last = 0;
int cost = 0, highest_cost = 0;
int range_start = 0, range_end = 0;
for ( int i=0; i<nTags; i++) {
mutlib_tag_t* pTag = MutScanGetTag( &ms, i );
cost += 4;
if (i != range_start)
cost -= pTag->Position[0] - last;
printf("Tag %d at %d, cost = %d\n", i, pTag->Position[0], cost);
if (highest_cost <= cost) {
highest_cost = cost;
range_end = i;
}
if ((cost < highest_cost-10 || cost < 0) && highest_cost > 10) {
printf(" Tag %d..%d highest_cost=%d\n",
range_start, range_end, highest_cost);
i = range_end;
range_start = i+1;
highest_cost = cost = 0;
}
last = pTag->Position[0];
}
if (highest_cost > 10) {
printf(" Tag %d..%d highest_cost=%d\n",
range_start, range_end, highest_cost);
}
#endif
#if 0
// Adjust left end
int cost = -5, last = cov_start;
puts("==Left==");
for( int i=0; i<nTags; i++ ) {
mutlib_tag_t* pTag = MutScanGetTag( &ms, i );
assert(pTag != NULL);
if (std::strcmp(pTag->Type,"MCOV") == 0)
continue;
cost += pTag->Position[0] - last;
printf("Tag at %d cost=%d\n", pTag->Position[0], cost);
if (cost <= 0) {
printf("Reject tag at %d\n", pTag->Position[0]);
*pTag->Type = 0;
cost -= 1;
}
last = pTag->Position[0];
}
// Adjust right end
cost = -5;
last = cov_end;
puts("==Right==");
for( int i=nTags-1; i>=0; i-- ) {
mutlib_tag_t* pTag = MutScanGetTag( &ms, i );
assert(pTag);
if (std::strcmp(pTag->Type,"MCOV") == 0)
continue;
cost += last - pTag->Position[0];
printf("Tag at %d cost=%d\n", pTag->Position[0], cost);
if (cost <= 0) {
printf("Reject tag at %d\n", pTag->Position[0]);
*pTag->Type = 0;
cost -= 2;
}
last = pTag->Position[0];
}
#endif
}
//-----------
// Tracediff
//-----------
int main( int argc, char* argv[] )
{
int n;
char* p;
mutscan_t ms;
mutlib_strand_t nStrand;
StringList FileList;
int nInputClipL;
int nInputClipR;
char pBuffer[BUFSIZE];
char pFileOfFiles[BUFSIZE] = { 0 };
Exp_info* pExpFile = 0;
Read* pInputTrace = 0;
int nRefClipL[2] = { -1, -1 };
int nRefClipR[2] = { -1, -1 };
Read* pRefTrace[2] = { 0, 0 };
char* pRefSequence[2] = { 0, 0 };
bool bDebug = false;
bool bQuiet = false;
bool bComplementTags = false;
double nNoiseThreshold = -1.0;
double nAlignFailThreshold = -1.0;
double nSearchWindowSize = -1.0;
double nPeakDropThresholdUpper = -1.0;
double nPeakDropThresholdLower = -1.0;
double nHeterozygoteSNRThreshold = -1.0;
int proximityThreshold = 7;
MutScanInit( &ms );
try
{
// Request traces to be used in preference to experiment files
read_experiment_redirect(0);
// Quick check of input
if( argc < 2 )
{
PrintBanner();
PrintUsage( ms );
return -1;
}
// Parse the command line options
for( n=1; n<argc; n++ )
{
// All options done?
if( argv[n][0] != '-' )
break;
// Copy switch contents to buffer
char c;
int k = 2;
while( c=argv[n][k] )
{
pBuffer[k-2] = c;
k++;
}
pBuffer[k-2] = 0;
// Process each option
switch( argv[n][1] )
{
case 'a':
// Alignment failure threshold
nAlignFailThreshold = std::atof( pBuffer );
break;
case 'c':
// Complement reverse strand base tags
bComplementTags = true;
break;
case 'f':
// File of filenames
std::strcpy( pFileOfFiles, pBuffer );
break;
case 'h':
// Heterozygote SNR ratio threshold
nHeterozygoteSNRThreshold = std::atof( pBuffer );
break;
case 'l':
// Lower peakdrop threshold
nPeakDropThresholdLower = std::atof( pBuffer );
break;
case 'n':
// Noise window
nNoiseThreshold = std::atof( pBuffer );
break;
case 'q':
// Quiet mode on
bQuiet = true;
break;
case 'u':
// Upper peakdrop threshold
nPeakDropThresholdUpper = std::atof( pBuffer );
break;
case 'w':
// Search window size
nSearchWindowSize = std::atof( pBuffer );
break;
case 't':
// proximity threshold
proximityThreshold = std::atoi( pBuffer );
break;
case 'z':
// Debug mode
bDebug = true;
break;
default:
// Unknown switch
std::fprintf( stderr, "Unknown option %s, ignoring.", argv[n] );
std::fflush( stderr );
break;
}
}
// Show banner
if( !bQuiet )
PrintBanner();
// Loop for MSW debugger attachment purposes
while( bDebug );
// Read contents of file-of-filenames if supplied
if( pFileOfFiles[0] )
{
std::FILE* pFOFN = std::fopen( pFileOfFiles, "rt" );
if( pFOFN )
{
while( std::fgets(pBuffer,BUFSIZE,pFOFN) )
{
p = std::strrchr(pBuffer,'\n');
if(p) *p = 0;
p = std::strrchr(pBuffer,'\r');
if(p) *p = 0;
FileList.Append( pBuffer );
}
std::fclose( pFOFN );
}
else
{
std::fprintf( stderr, "Unable to open file of filenames %s, skipping.\n", pFileOfFiles );
std::fflush( stderr );
}
}
// Add any experiment files on the command line
for( n=n; n<argc; n++ )
FileList.Append( argv[n] );
if( FileList.Length() < 1 )
{
PrintUsage( ms );
return -1;
}
// Set parameters if specified, otherwise accept defaults
if( nAlignFailThreshold > 0.0 )
MutScanSetParameter( &ms, MUTSCAN_PARAMETER_ALIGNFAIL_THRESHOLD, nAlignFailThreshold );
if( nHeterozygoteSNRThreshold > 0.0 )
MutScanSetParameter( &ms, MUTSCAN_PARAMETER_HETSNR_THRESHOLD, nHeterozygoteSNRThreshold );
if( nNoiseThreshold > 0.0 )
MutScanSetParameter( &ms, MUTSCAN_PARAMETER_NOISE_THRESHOLD, nNoiseThreshold );
if( nSearchWindowSize > 0.0 )
MutScanSetParameter( &ms, MUTSCAN_PARAMETER_SEARCH_WINDOW, nSearchWindowSize );
if( nPeakDropThresholdLower > 0.0 )
MutScanSetParameter( &ms, MUTSCAN_PARAMETER_PEAKDROP_LOWER, nPeakDropThresholdLower );
if( nPeakDropThresholdUpper > 0.0 )
MutScanSetParameter( &ms, MUTSCAN_PARAMETER_PEAKDROP_UPPER, nPeakDropThresholdUpper );
if( bComplementTags == true )
MutScanSetParameter( &ms, MUTSCAN_PARAMETER_COMPLEMENT_TAGS, 1.0 );
// File processing loop
for( n=0; n<FileList.Length(); n++ )
{
// Cleanup from previous iteration
if( pExpFile )
{
exp_destroy_info(pExpFile);
pExpFile = 0;
}
if( pInputTrace )
{
read_deallocate(pInputTrace);
pInputTrace = 0;
}
p = (n==0) ? FileList.First() : FileList.Next();
// Tell user what we're doing
if( !bQuiet )
{
std::fprintf( stdout, "Scanning: %s\n", p );
std::fflush( stdout );
}
// Open experiment file
pExpFile = exp_read_info( p );
if( !pExpFile )
{
std::fprintf( stderr, "Unable to open experiment file %s, skipping.\n", p );
std::fflush( stderr );
continue;
}
// Extract PR record, gives us strand direction
int PR = 0;
if( exp_get_int(pExpFile,EFLT_PR,&PR) )
{
std::fprintf( stderr, "Unable to read PR record from experiment file %s, assuming forward strand.\n", p );
std::fflush( stderr );
}
nStrand = ((PR==0) || (PR==1) || (PR==3)) ? MUTLIB_STRAND_FORWARD : MUTLIB_STRAND_REVERSE;
// If we don't have a reference trace/seq for this strand
if( !pRefTrace[nStrand] )
{
// Extract reference trace name WT
if( exp_get_str(pExpFile,EFLT_WT,pBuffer,BUFSIZE) )
{
std::fprintf( stderr, "Unable to read WT record from experiment file %s, skipping.\n", p );
std::fflush( stderr );
continue;
}
// Open reference trace file
MakeFullPath( p, pBuffer );
pRefTrace[nStrand] = read_reading( pBuffer, TT_ANY );
if( !pRefTrace[nStrand] )
{
std::fprintf( stderr, "Unable to open reference trace file %s, skipping.\n", pBuffer );
std::fflush( stderr );
continue;
}
// Extract reference clip points WL/WR
exp_get_int( pExpFile, EFLT_WL, &nRefClipL[nStrand] );
exp_get_int( pExpFile, EFLT_WR, &nRefClipR[nStrand] );
// If default clipping values used for reference
if( (nRefClipL[nStrand]<0) || (nRefClipR[nStrand]<0) )
{
// Construct the reference experiment file's name and attempt to open it
ReplaceExtension( pBuffer, ".exp" );
Exp_info* pRefExpFile = exp_read_info( pBuffer );
if( pRefExpFile )
{
// Extract the QL/QR records if they exist
if( nRefClipL[nStrand] < 0 )
exp_get_int( pRefExpFile, EFLT_QL, &nRefClipL[nStrand] );
if( nRefClipR[nStrand] < 0 )
exp_get_int( pRefExpFile, EFLT_QR, &nRefClipR[nStrand] );
exp_destroy_info(pRefExpFile);
}
// Otherwise use the entire reference trace
}
// Initialise the algorithm
MutScanSetReference( &ms, nStrand, pRefTrace[nStrand], nRefClipL[nStrand], nRefClipR[nStrand] );
}
// Extract LN record, gives us input trace file name
if( exp_get_str(pExpFile,EFLT_LN,pBuffer,BUFSIZE) )
{
std::fprintf( stderr, "Unable to read LN record from experiment file %s, skipping.\n", p );
std::fflush( stderr );
continue;
}
// Open input trace
MakeFullPath( p, pBuffer );
pInputTrace = read_reading( pBuffer, TT_ANY );
if( !pInputTrace )
{
std::fprintf( stderr, "Unable to open input trace file %s, skipping.\n", pBuffer );
std::fflush( stderr );
continue;
}
if ( !pInputTrace->NPoints ) {
std::fprintf(stderr, "No trace data found in trace file %s, skipping.\n", pBuffer);
continue;
}
// Extract input clip points QL/QR
nInputClipL = -1;
nInputClipR = -1;
exp_get_int( pExpFile, EFLT_QL, &nInputClipL );
exp_get_int( pExpFile, EFLT_QR, &nInputClipR );
// Initialise the algorithm
MutScanSetInput( &ms, nStrand, pInputTrace, nInputClipL, nInputClipR );
// Execute the algorithm
MutScanExecute( &ms );
if( MutScanGetResultCode(&ms) )
{
// Error
std::fprintf( stderr, "%s", MutScanGetResultString(&ms) );
std::fflush( stderr );
continue;
}
int nTags = MutScanGetTagCount( &ms );
// Find the rightmost tag position for this trace
int nRightmostTag = -1;
for( int i=0; i<nTags; i++ )
{
mutlib_tag_t* pTag = MutScanGetTag( &ms, i );
if( pTag->Position[0] > nRightmostTag )
nRightmostTag = pTag->Position[0];
}
// If mutation tag is beyond the right clip point (as in the case of
// insertions/deletions), we adjust QR so that it's visible in gap4.
if( (nTags>0) && (nInputClipR<=nRightmostTag) )
{
std::sprintf( pBuffer, "%d", nRightmostTag+1 );
exp_put_str( pExpFile, EFLT_QR, pBuffer, std::strlen(pBuffer) );
}
// Filter clusters of tags too close to the end of MCOV
filter_tags( ms, nTags, proximityThreshold );
// Output results
for( int i=0; i<nTags; i++ )
{
// Write mutation tags to experiment file & stdout
mutlib_tag_t* pTag = MutScanGetTag( &ms, i );
assert(pTag != NULL);
if (!*pTag->Type)
continue;
bool bCoverageTag = std::strcmp(pTag->Type,"MCOV") == 0;
char sc = (pTag->Strand==MUTLIB_STRAND_FORWARD) ? '+' : '-';
if( bCoverageTag )
{
std::sprintf( pBuffer, "%s %c %d..%d", pTag->Type, sc, pTag->Position[0], pTag->Position[1] );
if( !bQuiet )
{
std::fprintf( stdout, "%s\n", pBuffer );
std::fflush( stdout );
}
}
else
{
std::sprintf( pBuffer, "%s %c %d..%d\n%s", pTag->Type, sc, pTag->Position[0], pTag->Position[1], pTag->Comment );
if( !bQuiet )
{
std::fprintf( stdout, "%s %5d %s\n", pTag->Type, pTag->Position[0], pTag->Comment );
std::fflush( stdout );
}
}
exp_put_str(pExpFile, EFLT_TG, pBuffer, std::strlen(pBuffer) );
}
}
}
catch( std::bad_alloc& )
{
// Out of memory
std::fprintf( stderr, "%s", "Not enough memory to complete the operation.\n" );
std::fflush( stderr );
}
catch(...)
{
// Print error message
std::fprintf( stderr, "%s", "An unexpected fatal exception has occurred, please "
"report the details to staden-package@mrc-lmb.cam.ac.uk.\n" );
std::fflush( stderr );
}
// Cleanup & exit
if(pExpFile) exp_destroy_info(pExpFile);
if(pInputTrace) read_deallocate(pInputTrace);
if(pRefTrace[0]) read_deallocate(pRefTrace[0]);
if(pRefTrace[1]) read_deallocate(pRefTrace[1]);
MutScanDestroy( &ms );
return 0;
}
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