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  • Comparing mapped reads

    I have used Tophat to map reads to a reference sequences. I would like to identify reads that mapped to multiple sequences. Does anyone know how this can be accomplished?

    E.g. I would like to be able to say, read 1 mapped to reference sequences 1, 3, 23, and 67.

    The above would be ideal... but if that is not possible then something like reference sequence 3 had reads 1, 19, 394, 39291, etc. map to it.

    Any ideas? I feel like this is probably pretty doable I am just a rookie though.

  • #2
    Look at the NH:i flag at the end of the line. This indicates how many loci the read mapped to. The third column shows which reference sequence the alignment is to.

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    • #3
      TO GET CLASSIFICATION OF MAPPED READS
      "samtools view sorted.bam | awk '{print $2}' | sort | uniq -c"
      #THIS GIVES A RESULT
      83839545 0
      83824316 16
      6755765 256
      6850132 272

      HERE 0, 16, 256, 272 ARE TOPHAT FLAGS TO UNDERSTAND THEIR MEANING GO TO:http://picard.sourceforge.net/explain-flags.html

      GIVES THE COUNT OF NUMBER OF UNIQUE READS MAPPED

      "samtools view t24hpf_sorted.bam | awk '{print $1}' | sort | uniq -c"

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