BWA - samtools problem

[genelab]

Hello everyone

There are some topics concerning BWA and Samtools, but i still can't find the reasons of the trouble.

Here is what I count

bwa index -a bwtsw human.fasta
bwa aln human.fasta read.fq > aln_sa.sai
bwa samse human.fasta aln_sa.sai read.fq > aln.sam
samtools view -b -h -S -T human.fasta aln.sam > aln.bam
samtools sort aln.bam aln.sort
samtools index aln.sort.bam

when i finished these steps and get the sorted bam file, i then run

java -jar ViewSam.jar I=aln.sort.bam

it got the following error message:


Exception in thread "main" java.lang.RuntimeException: SAM validation error: ERROR: Record 1, Read name 1_R9DycrMpuN1, MRNM should not be set for unpaired read.
at net.sf.samtools.SAMUtils.processValidationErrors(SAMUtils.java:334)
at net.sf.samtools.BAMFileReader$BAMFileIterator.advance(BAMFileReader.java:444)
at net.sf.samtools.BAMFileReader$BAMFileIterator.<init>(BAMFileReader.java:408)
at net.sf.samtools.BAMFileReader$BAMFileIterator.<init>(BAMFileReader.java:398)
at net.sf.samtools.BAMFileReader.getIterator(BAMFileReader.java:201)
at net.sf.samtools.SAMFileReader.iterator(SAMFileReader.java:283)
at net.sf.samtools.SAMFileReader.iterator(SAMFileReader.java:38)
at net.sf.picard.sam.ViewSam.doWork(ViewSam.java:68)
at net.sf.picard.cmdline.CommandLineProgram.instanceMain(CommandLineProgram.java:150)
at net.sf.picard.sam.ViewSam.main(ViewSam.java:58)

how can i fix this?
Thanks!!!

[lh3]

Give the first line of your alignment, by samtools.

[genelab]

I have found one way to figure out the problem

I used the function "setValidationStringency" in samtools


thx!