Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • wintergreen36
    Member
    • Jul 2014
    • 19

    Demultiplexing with last three bases

    What's the possibility of using the last three bases in the index for demultiplexing the HiSeq 2500 data using bcl2fstq

    Let me say index is GCATCG
    I wish to de multiplex using the last three bases TCG only ,
    The quality scores for index read were bad; it's 40 %

    how to give a variable for serving the above said purposes.
  • GenoMax
    Senior Member
    • Feb 2008
    • 7142

    #2
    You can check (even by eye) to see if that is possible by looking at the indexes you actually used. If two or more overlap (in last three bases that you want to use) then you won't be able to demultiplex.

    Comment

    • wintergreen36
      Member
      • Jul 2014
      • 19

      #3
      yaya am aware of that , let me say

      every lane got 6 samples without overlapping the last three bases ,
      if its a simple 50 SR run , how to write a varibale to mask the first three bases and demultiplexing using only last three bases

      Comment

      • GenoMax
        Senior Member
        • Feb 2008
        • 7142

        #4
        --use-bases-mask nnnYYY.
        Last edited by GenoMax; 08-12-2014, 05:10 AM.

        Comment

        • wintergreen36
          Member
          • Jul 2014
          • 19

          #5
          Below command line includes the path too see the last line

          /usr/local/bin/configureBclToFastq.pl --input-dir /media/vanessa/Sandor_1/20140408SANDORSR50/140408_700417L_0315_BC4C4GACXX/Data/Intensities/BaseCalls --output-dir /media/vanessa/Sandor_1/Unaligned --sample-sheet /media/vanessa/Sandor_1/20140408SANDORSR50/140408_700417L_0315_BC4C4GACXX/Data/Intensities/BaseCalls/SampleSheet.csv --USE_BASES nnnYYY
          will it work like this ?

          Comment

          • GenoMax
            Senior Member
            • Feb 2008
            • 7142

            #6
            Please see the updated command option in post #4. Should be --use-bases-mask nnnYYY.

            Forgot about Read 1. Rick has provided the correct full syntax below.
            Last edited by GenoMax; 08-12-2014, 08:27 AM.

            Comment

            • westerman
              Rick Westerman
              • Jun 2008
              • 1104

              #7
              GenoMax undoubtedly has the syntax correct for a single-end demultiplex [I mostly do paired-end] but it is possible that you may need the full specification of

              use-bases-mask=Y*,InnnYYY

              or maybe

              use-bases-mask=Y*,InnnYYYn

              Comment

              • wintergreen36
                Member
                • Jul 2014
                • 19

                #8
                Thank You very much @ Genomax @ westerman

                Comment

                • wintergreen36
                  Member
                  • Jul 2014
                  • 19

                  #9
                  @ hi genomax and westerman , which version of ubuntu u experienced best to installa and run casava and bcl2fastq :-)

                  Comment

                  • GenoMax
                    Senior Member
                    • Feb 2008
                    • 7142

                    #10
                    Originally posted by wintergreen36 View Post
                    @ hi genomax and westerman , which version of ubuntu u experienced best to installa and run casava and bcl2fastq :-)
                    Exact version of ubuntu should not matter as long as the software works.

                    Comment

                    Latest Articles

                    Collapse

                    • SEQadmin2
                      From Collection to Sequencing: Why Sample Preparation and Preservation Define Sequencing Data
                      by SEQadmin2


                      Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.


                      The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
                      ...
                      06-02-2026, 10:05 AM
                    • SEQadmin2
                      Single-Cell Sequencing at an Inflection Point: Early Impacts of New Platforms and Emerging Trends
                      by SEQadmin2


                      With the launch of new single-cell sequencing platforms in 2026, the field stands at an exciting inflection point. This article surveys the most impactful advances in the field and discusses how they’re reshaping research in cancer, immunology, and beyond.


                      Introduction

                      Single-cell sequencing technologies have undergone remarkable advances over the past decade, transitioning from low-throughput experimental approaches to highly scalable platforms capable of...
                      05-22-2026, 06:42 AM
                    • SEQadmin2
                      Environmental Genomics in the Age of NGS: From Microbes to Conservation Strategies
                      by SEQadmin2

                      Studying ecosystems means dealing with complex, multi-species communities that are hard to observe at scale. This complexity, however, hides many important questions to be answered, from how biogeochemical cycles work and how climate change can affect species distribution to how conservation strategies can work best.


                      Genomics, particularly since the expansion of NGS, has transformed ecosystem ecology. By sequencing environmental DNA, we can now assess biodiversity without direct...
                      05-06-2026, 09:04 AM

                    ad_right_rmr

                    Collapse

                    News

                    Collapse

                    Topics Statistics Last Post
                    Started by SEQadmin2, Today, 08:59 AM
                    0 responses
                    4 views
                    0 reactions
                    Last Post SEQadmin2  
                    Started by SEQadmin2, 06-02-2026, 12:03 PM
                    0 responses
                    21 views
                    0 reactions
                    Last Post SEQadmin2  
                    Started by SEQadmin2, 06-02-2026, 11:40 AM
                    0 responses
                    14 views
                    0 reactions
                    Last Post SEQadmin2  
                    Started by SEQadmin2, 05-28-2026, 11:40 AM
                    0 responses
                    29 views
                    0 reactions
                    Last Post SEQadmin2  
                    Working...