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  • wintergreen36
    Member
    • Jul 2014
    • 19

    #1

    demultiplexing Nextera Rapied Capture exome data on bcl2fastq 1.8.3

    I am using bcl2fastq 1.8.3 on ubuntu 14.04 LTS.

    What's the best variable can be given for demultiplexing Nextera Rapied Capture exome data

    details

    Next era indice - 8 bases
    RUN TYPE - read 1(100 cycles), read 2(8cycles) , read 3(100cycles),
  • GenoMax
    Senior Member
    • Feb 2008
    • 7142

    #2
    Originally posted by wintergreen36 View Post
    What's the best variable can be given for demultiplexing Nextera Rapied Capture exome data
    Not sure I understand the question. You will have to clarify further.

    Comment

    • wintergreen36
      Member
      • Jul 2014
      • 19

      #3
      I tried to demultiplex nexera exome data with a normal command , it gave an error like barcode lenght including delimter is 7

      so I gave varibale like Y100n,I8,Y100n is this correct ?

      Comment

      • GenoMax
        Senior Member
        • Feb 2008
        • 7142

        #4
        Use bases mask is generally determined automatically from the RunInfo.xml file.

        You can use Y100n,I7n,Y100n or edit your samplesheet and make the tags 7 bp long.

        Comment

        • wintergreen36
          Member
          • Jul 2014
          • 19

          #5
          @genomax is there any wrong if we declared index as ,I8

          Comment

          • GenoMax
            Senior Member
            • Feb 2008
            • 7142

            #6
            Last base in the read (even for tags) is generally not considered (because it lacks phasing information). So when bcl2fastq/CASAVA demultiplexes a run it will use n-1 cycles for demultiplexing. You can use an appropriate "--use-bases-mask" or adjust your samplesheet when setting up a demultiplexing run. If you need 8 bases on the tag reads then set sequencing run for 9 cycles.

            Comment

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