Hi Shanrong,

You can try one or all of these:

 * Add option VALIDATION_STRINGENCY=LENIENT or
   VALIDATION_STRINGENCY=SILENT to the SamToFastq command line.
 * Investigate the way your BAM file was created, to try to understand
   why an invalid bin field was put into the file.
 * Convert BAM to SAM and then back to BAM again.  Unfortunately, this
   is the only way I know of to fix the bin fields in a BAM file.

-Alec


On 7/1/14, 9:08 PM, Zhao, Shanrong wrote:

I have difficulty in doing this simple thing using PICARD. I tried the latest version as well as an older version 1.110, but got the same error message below. I simply don't how to walk around this issue.

[Tue Jul 01 20:22:44 EDT 2014] net.sf.picard.sam.SamToFastq INPUT=LP6005799-DNA_A02.bam FASTQ=LP6005799-DNA_A02_1.fastq SECOND_END_FASTQ=LP6005799-DNA_A02_2.fastq UNPAIRED_FASTQ=LP6005799-DNA_A02.unpaired.fastq OUTPUT_PER_RG=false RE_REVERSE=true INTERLEAVE=false INCLUDE_NON_PF_READS=false READ1_TRIM=0 READ2_TRIM=0 INCLUDE_NON_PRIMARY_ALIGNMENTS=false VERBOSITY=INFO QUIET=false VALIDATION_STRINGENCY=STRICT COMPRESSION_LEVEL=5 MAX_RECORDS_IN_RAM=500000 CREATE_INDEX=false CREATE_MD5_FILE=false

[Tue Jul 01 20:22:44 EDT 2014] Executing as [email protected] on Linux 2.6.32-358.11.1.el6.x86_64 amd64; OpenJDK 64-Bit Server VM 1.7.0_25-mockbuild_2013_06_26_07_26-b00; Picard version: 1.110(1752) IntelDeflater

INFO 2014-07-01 20:23:04 SamToFastq Processed 1,000,000 records. Elapsed time: 00:00:19s. Time for last 1,000,000: 18s. Last read position: chrM:13,788

[Tue Jul 01 20:23:06 EDT 2014] net.sf.picard.sam.SamToFastq done. Elapsed time: 0.38 minutes.

Runtime.totalMemory()=3668639744

To get help, see http://picard.sourceforge.net/index.shtml#GettingHelp

Exception in thread "main" net.sf.samtools.SAMFormatException: SAM validation error: ERROR: Record 1179125, Read name C30BLACXX_0:5:1313:824748:0, bin field of BAM record does not equal value computed based on alignment start and end, and length of sequence to which read is aligned

at net.sf.samtools.SAMUtils.processValidationErrors(SAMUtils.java:449)

at net.sf.samtools.BAMFileReader$BAMFileIterator.advance(BAMFileReader.java:621)

at net.sf.samtools.BAMFileReader$BAMFileIterator.next(BAMFileReader.java:606)

at net.sf.samtools.BAMFileReader$BAMFileIterator.next(BAMFileReader.java:576)

at net.sf.samtools.SAMFileReader$AssertableIterator.next(SAMFileReader.java:774)

at net.sf.samtools.SAMFileReader$AssertableIterator.next(SAMFileReader.java:752)

        at net.sf.picard.sam.SamToFastq.doWork(SamToFastq.java:139)

at net.sf.picard.cmdline.CommandLineProgram.instanceMain(CommandLineProgram.java:179)

        at net.sf.picard.sam.SamToFastq.main(SamToFastq.java:127)



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Turn processes into business applications with Bonita BPM Community Edition
Quickly connect people, data, and systems into organized workflows
Winner of BOSSIE, CODIE, OW2 and Gartner awards
http://p.sf.net/sfu/Bonitasoft
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