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Question: MarkDuplicates output file in GATK pipeline
1
gravatar for gprashant17
4 weeks ago by
gprashant17 • 60
gprashant17 • 60 wrote:

I have used GATK's MarkDuplicates on a BAM file I obtained after alignment, which resulted in another file marked_duplicates.bam. So should I proceed with this marked_duplicates.bam file for analysis (converting to VCF), or this is just a file containing duplicates? In the latter case, is it possible to obtain a BAM file, with all the duplicates removed?
sequencing rna-seq bam alignment gatk • 92 views
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modified 4 weeks ago by Pierre Lindenbaum ♦ 121k • written 4 weeks ago by gprashant17 • 60
1
gravatar for Pierre Lindenbaum
4 weeks ago by
Pierre Lindenbaum ♦ 121k
France/Nantes/Institut du Thorax - INSERM UMR1087
Pierre Lindenbaum ♦ 121k wrote:

So should I proceed with this marked_duplicates.bam file for analysis (converting to VCF),

yes. As a proof, test both files wth samtools flagstats

is it possible to obtain a BAM file, with all the duplicates removed?

in the manual : https://software.broadinstitute.org/gatk/documentation/tooldocs/4.0.4.0/picard_sam_markduplicates_MarkDuplicates.php#--REMOVE_DUPLICATES

--REMOVE_DUPLICATES / NA

If true do not write duplicates to the output file instead of writing them with appropriate flags set.
ADD COMMENT • link written 4 weeks ago by Pierre Lindenbaum ♦ 121k

So if I did not use --REMOVE_DUPLICATES, the duplicate reads will still be present in the marked_duplicates.bam but they would have been flagged as duplicates right?

ADD REPLY • link written 4 weeks ago by gprashant17 • 60

So if I did not use --REMOVE_DUPLICATES, the duplicate reads will still be present in the marked_duplicates.bam but they would have been flagged as duplicates right?

yes

ADD REPLY • link written 4 weeks ago by Pierre Lindenbaum ♦ 121k
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