20 April 2010

Reading/Writing SAM/BAM files with the picard API for java.

I'm currently playing with the BAM/SAM Java API available in the picard package. This API can be used for creating new programs that read and write SAM files, the generic format for storing large nucleotide sequence alignments. Both SAM text format and SAM binary (BAM) format are supported. The source code comes with a simple example showing how to read and write a SAM/BAM file.

(...)
public void convertReadNamesToUpperCase(
final File inputSamOrBamFile,
final File outputSamOrBamFile
)
{
final SAMFileReader inputSam = new SAMFileReader(inputSamOrBamFile);
final SAMFileWriter outputSam = new SAMFileWriterFactory().makeSAMOrBAMWriter(inputSam.getFileHeader(),
true, outputSamOrBamFile);

for (final SAMRecord samRecord : inputSam) {
// Convert read name to upper case.
samRecord.setReadName(samRecord.getReadName().toUpperCase());
outputSam.addAlignment(samRecord);
}
outputSam.close();
inputSam.close();
}
(...)
toay, I've played with this SAM API to build two simple java tools:

SbamGrep

SbamGrep is available at http://code.google.com/p/code915/wiki/SbamGrep. It filters a SAM/BAM file using the SAM flags.

Option

 -o (filename-out) or default is stdout SAM
-v inverse selection
-f [flag] add bam/sam flag for filtering. multiple separeted with a comma. One of:
READ_PAIRED or 1 or 0x1
PROPER_PAIR_ or 2 or 0x2
READ_UNMAPPED or 4 or 0x4
MATE_UNMAPPED or 8 or 0x8
READ_STRAND or 16 or 0x10
MATE_STRAND or 32 or 0x20
FIRST_OF_PAIR or 64 or 0x40
SECOND_OF_PAIR or 128 or 0x80
NOT_PRIMARY_ALIGNMENT or 256 or 0x100
READ_FAILS_VENDOR_QUALITY_CHECK or 512 or 0x200
DUPLICATE_READ or 1024 or 0x400

Example

The following command lists all the reads having a flag (READ_PAIRED and READ_UNMAPPED and MATE_UNMAPPED and FIRST_OF_PAIR):
java -cp sam-1.16.jar:dist/sbamgrep.jar fr.inserm.umr915.sbamtools.SbamGrep -f 0x4,0x1,0x8,0x40 file.bam
Result (masked):
@HD VN:1.0 SO:unsorted
@SQ SN:chrT LN:349250
IL_XXXX1 77 * 0 0 * * 0 0 XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX (94**0-)*7=0688855555522@86;;;5;6:;63:4?-622647..-.5.%
IL_XXXX2 77 * 0 0 * * 0 0 XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX (9*+*2396,@5+5:@@@;;5)50)6960684;58;86*5102)0*+8:*137;
IL_XXXX3 77 * 0 0 * * 0 0 XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX (/999-00328:88984@@=8??@@:-66,;8;5;6+;255,1;788883676'
IL_XXXX4 77 * 0 0 * * 0 0 XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX (916928.82@@50854;33222224;@25?5522;5=;;858888555*0666
IL_XXXX5 77 * 0 0 * * 0 0 XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX (9*4*5-**32989+::@82;;853+39;80.53)-)79)..'55.8988*200
IL_XXXX6 77 * 0 0 * * 0 0 XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX (*+**,14265;@@??@8?9@@@5@488?8666260.)-*9;;;88:8'05418
IL_XXXX7 77 * 0 0 * * 0 0 XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX (9136242-2@@@;96.888@@@@80$585882623.':**+3*03137..--.
IL_XXXX8 77 * 0 0 * * 0 0 XXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXXX (/89255855@88?585557..())@@@;5552286526755@@5888..3;/$
(...)

SbamStats

The second tool, SbamStats is available at http://code.google.com/p/code915/wiki/SbamStats. It provides a simple report about all the SAM flags used in one or more files.

Example

:
java -cp sam-1.16.jar:dist/sbamstats.jar fr.inserm.umr915.sbamtools.SBamStats file.bam
Output:
READ_PAIRED:0x1|READ_STRAND:0x10|MATE_STRAND:0x20|FIRST_OF_PAIR:0x40 10827
READ_PAIRED:0x1|READ_STRAND:0x10|MATE_STRAND:0x20|SECOND_OF_PAIR:0x80 10827
READ_PAIRED:0x1|FIRST_OF_PAIR:0x40 13951
READ_PAIRED:0x1|SECOND_OF_PAIR:0x80 13951
READ_PAIRED:0x1|MATE_STRAND:0x20|FIRST_OF_PAIR:0x40 23846
READ_PAIRED:0x1|READ_STRAND:0x10|SECOND_OF_PAIR:0x80 23846
READ_PAIRED:0x1|MATE_STRAND:0x20|SECOND_OF_PAIR:0x80 33606
READ_PAIRED:0x1|READ_STRAND:0x10|FIRST_OF_PAIR:0x40 33606
READ_PAIRED:0x1|MATE_UNMAPPED:0x8|READ_STRAND:0x10|SECOND_OF_PAIR:0x80 143090
READ_PAIRED:0x1|READ_UNMAPPED:0x4|READ_STRAND:0x10|MATE_STRAND:0x20|FIRST_OF_PAIR:0x40 143090
READ_PAIRED:0x1|READ_UNMAPPED:0x4|SECOND_OF_PAIR:0x80 161781
READ_PAIRED:0x1|MATE_UNMAPPED:0x8|FIRST_OF_PAIR:0x40 161781
READ_PAIRED:0x1|MATE_UNMAPPED:0x8|SECOND_OF_PAIR:0x80 174429
READ_PAIRED:0x1|READ_UNMAPPED:0x4|FIRST_OF_PAIR:0x40 174429
READ_PAIRED:0x1|MATE_UNMAPPED:0x8|READ_STRAND:0x10|FIRST_OF_PAIR:0x40 176219
READ_PAIRED:0x1|READ_UNMAPPED:0x4|READ_STRAND:0x10|MATE_STRAND:0x20|SECOND_OF_PAIR:0x80 176219
READ_PAIRED:0x1|PROPER_PAIR_:0x2|MATE_STRAND:0x20|FIRST_OF_PAIR:0x40 915161
READ_PAIRED:0x1|PROPER_PAIR_:0x2|READ_STRAND:0x10|SECOND_OF_PAIR:0x80 915161
READ_PAIRED:0x1|PROPER_PAIR_:0x2|MATE_STRAND:0x20|SECOND_OF_PAIR:0x80 2623345
READ_PAIRED:0x1|PROPER_PAIR_:0x2|READ_STRAND:0x10|FIRST_OF_PAIR:0x40 2623345
READ_PAIRED:0x1|READ_UNMAPPED:0x4|MATE_UNMAPPED:0x8|SECOND_OF_PAIR:0x80 26232123
READ_PAIRED:0x1|READ_UNMAPPED:0x4|MATE_UNMAPPED:0x8|FIRST_OF_PAIR:0x40 26232123



That's it !
Pierre

1 comment:

Chaz said...

I just downloaded both of these scripts and they have been pretty cool so far. I am presenting in lab meeting about the summary one today. Thanks!