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  • Picard, Markduplicates

    We use Picard/CleanSama.jar:

    java -Xmx4g -jar $Picard_jarFolder/CleanSam.jar INPUT=$sample OUTPUT=$clean_picard VALIDATION_STRINGENCY=LENIENT 2>$log_clean_picard"

    Later we use fixmate a sort by samtools.
    but when we use the next step, to mark duplicates:

    java -Xmx2g -jar $Picard_jarFolder/MarkDuplicates.jar INPUT=$sorted1_samtools_bam OUTPUT=$sorted_picard_marked_bam TMP_DIR=./ METRICS_FILE=$log_duplicate_metrics REMOVE_DUPLICATES=false ASSUME_SORTED=true VALIDATION_STRINGENCY=SILENT CREATE_INDEX=true 2>$log_picard_marked_bam"

    We have the next problem and we dont know why, if we had used CleanSam (which it is supposed to Cleans the provided SAM/BAM, soft-clipping beyond-end-of-reference alignments and setting MAPQ to 0 for unmapped reads)

    [Thu Nov 13 12:43:12 CET 2014] net.sf.picard.sam.MarkDuplicates done. Elapsed time: 10,13 minutes.
    Runtime.totalMemory()=1691418624
    Exception in thread "main" net.sf.samtools.SAMException: Exception when processing alignment for BAM index HS12_10750:8:2310:1400:98064#42 1/2 75b unmapped read.
    at net.sf.samtools.BAMFileWriter.writeAlignment(BAMFileWriter.java:112)
    at net.sf.samtools.SAMFileWriterImpl.addAlignment(SAMFileWriterImpl.java:168)
    at net.sf.picard.sam.MarkDuplicates.doWork(MarkDuplicates.java:208)
    at net.sf.picard.cmdline.CommandLineProgram.instanceMain(CommandLineProgram.java:177)
    at net.sf.picard.sam.MarkDuplicates.main(MarkDuplicates.java:106)
    Caused by: net.sf.samtools.SAMException: Exception creating BAM index for record HS12_10750:8:2310:1400:98064#42 1/2 75b unmapped read.
    at net.sf.samtools.BAMIndexer.processAlignment(BAMIndexer.java:94)
    at net.sf.samtools.BAMFileWriter.writeAlignment(BAMFileWriter.java:109)
    ... 4 more
    Caused by: java.lang.ArrayIndexOutOfBoundsException: 8304
    at net.sf.samtools.BAMIndexer$BAMIndexBuilder.processAlignment(BAMIndexer.java:219)
    at net.sf.samtools.BAMIndexer.processAlignment(BAMIndexer.java:92)
    ... 5 more

    Previously, we have not reported this problem.

    Thanks.

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