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  • filter secondary reads in bam with pysam

    I am new to python and trying to learn. The below is an attempt to filter out secondary reads in a all bam files in a directory using pysam. I ran mark duplicates and got an error on several bam files due to secondary alignments. I added comments to each line to illustrate my thought process. Thank you .

    The specific mark duplicates error:

    Value was put into PairInfoMap more than once

    Code:
     
        #! /usr/bin/python  ## call python script
        import sys ## import python system functions
        import pysam  ## import module
    
        bam = pysam.AlignmentFile(.bam, "rb")  ## open bam and read
        click.echo("Reading BAM file")  ## output message
        for read in bam.fetch():
            if read.is_secondary=true  ## not the primary alignment
                continue
            if read.has_tag('AS') and read.has_tag('XS') ## look for Primary and Secondary Alignment Score tag
                AS = read.get_tag('AS')  ## read and store AS value
                XS = read.get_tag('XS')  ## read and store XS value
                     if AS >= XS  ## Alignment score greater then or equal to XS (these will be printed)
                bam.write(read)  ## open bam and only print primary alignments
                     continue  ## process next line
        bam.close()  ## end processing
    another option
    Code:
    #! /usr/bin/python  ## call python script
    import sys ## import python system functions
    import pysam  ## import module
    
    bam = pysam.AlignmentFile(bam, "rb")  ## open bam and read
    click.echo("Reading BAM file")  ## output message
    	for read in bam.fetch():  ## start loop and iterate over each bam
    		if read.is_secondary:  ## not the primary alignment
    			continue
    		if read.has_tag('AS') and read.has_tag('XS') ## look for Primary and Secondary Alignment Score tag
    			AS = read.get_tag('AS')  ## read and store AS value
    			XS = read.get_tag('XS')  ## read and store XS value
    				if AS >= XS  ## Alignment score greater then or equal to XS (these will be printed)
    		read.write(read)  ## only print primary alignments
    		continue
    bam.close()  ## end processing
    Last edited by cmccabe; 04-17-2019, 04:44 AM. Reason: added option 2

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