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  • illuminaGA
    Member
    • Dec 2012
    • 71

    #1

    Is this a overclustering?

    Dear All

    I am currently at 18 cycle of 101+7+101 PE running.

    I loaded 12pM sample and Phix(Lane4).

    The intensity is ok in the first base. But from cycle 2,the intensity was dropping dramatically.

    I also found the Base T thumbnail imagaes look strange.

    I attached some pictures here.

    What should I do now?

    Thank you so much?
    Attached Files
  • GenoMax
    Senior Member
    • Feb 2008
    • 7142

    #2
    Is this MiSeq or HiSeq? What is the cluster density number?

    You are not going to be able to do much if it is over-clustered. You will have to re-run.

    Comment

    • illuminaGA
      Member
      • Dec 2012
      • 71

      #3
      Here is other infomation.

      Only the bottom looks normal.
      Attached Files

      Comment

      • illuminaGA
        Member
        • Dec 2012
        • 71

        #4
        This is GAiix. cluster density is around 600.

        Thank you so much.

        Originally posted by GenoMax View Post
        Is this MiSeq or HiSeq? What is the cluster density number?

        You are not going to be able to do much if it is over-clustered. You will have to re-run.

        Comment

        • illuminaGA
          Member
          • Dec 2012
          • 71

          #5
          Originally posted by GenoMax View Post
          Is this MiSeq or HiSeq? What is the cluster density number?

          You are not going to be able to do much if it is over-clustered. You will have to re-run.
          In the previous run. Some time we got cluster density around 1000+/- 50. And got high quality data.

          But this time ,I don't konw why.
          Attached Files

          Comment

          • GenoMax
            Senior Member
            • Feb 2008
            • 7142

            #6
            Guess you should let the run finish and see what the data looks like. One hits a bad flowcell once in a while so this could just be a one time issue.

            Comment

            • illuminaGA
              Member
              • Dec 2012
              • 71

              #7
              Originally posted by GenoMax View Post
              Guess you should let the run finish and see what the data looks like. One hits a bad flowcell once in a while so this could just be a one time issue.
              Hi GenoMax, Thank you so much.

              I have to stop the running since the run will not perform well.Illumina Techsupport recommended checking the fluidics on the system by performing a maintenance wash and measuring the delivered volume by lane.
              I did it and fluidics were all good.
              Attached Files
              Last edited by illuminaGA; 07-01-2013, 09:08 AM.

              Comment

              • andibody
                Member
                • Mar 2008
                • 45

                #8
                The last picture reminds me of problems we had with the mode scrambler.

                Comment

                • illuminaGA
                  Member
                  • Dec 2012
                  • 71

                  #9
                  Thank you so much.

                  Illumina FSE will replace a new part for our GAiix.

                  Originally posted by andibody View Post
                  The last picture reminds me of problems we had with the mode scrambler.

                  Comment

                  • cdalgard
                    Member
                    • Apr 2011
                    • 12

                    #10
                    Did you ever figure out why you had a good first base report and then a significant decrease in intensities for Cycle 2?

                    Comment

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