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  • ECO
    --Site Admin--
    • Oct 2007
    • 1360

    #1

    MiSeq V2 Upgrade Information...

    There is some information in the original rumors thread...but I wanted to start a new thread to get some of the _actual_ post-V2-upgrade information out there (either from myself or others).

    We are still in the middle of our 2x250 phiX install runs on 2 of our MiSeqs, everything has gone really nicely so far. The hardware upgrade took basically 8 hours spread over two days with two FSE's working on it.

    Things I've noticed:

    • Flow cell part numbers are IDENTICAL between V1 and V2. Based on the part numbers on the external storage tube.
    • The V2 instruments are ~40 cycles ahead of the V1 instrument (which is running 2x151)
    • The intensity profiles are crazy different (see below)

    V1 intensity profile:



    V2 intensity profile:


    And %>Q30 looks similar but the higher quality in the V2 chemistry/imaging is evident comparing the quality at the same cycle...

    V1 %>Q30 (where at C150 it's at ~80%)


    V2 %>Q30 (where at C150 it's at 87%):


    Also, in the interest of benchmarking relative cluster densities, we ran the same dilution of the same phiX library (12.5pM according to the FAS) on one of our V1 instruments as well. He recommended not doing this because the observed V1 cluster density was expected to be higher than for V2.

    The two V2 instruments came up at 803 and 848...while the V1 run came up at 883. This is very close to our V1 experience, where we run 12pM to target 900K/mm^2, but it will be interesting to track if the moderately lower V2 cluster density is a real phenomenon.

    Anyway...looking great so far...stay tuned...I hope to run some real samples as soon as possible.
  • GW_OK
    Senior Member
    • Sep 2009
    • 411

    #2
    Wow, that is a crazy different intensity profile. And noisy Q30 plot, what's the deal with that?

    Comment

    • ECO
      --Site Admin--
      • Oct 2007
      • 1360

      #3
      Both Q30 plots look like that...the other instrument has a spot on one set of tiles so there is a bit of dropout:

      Comment

      • koadman
        Member
        • May 2010
        • 65

        #4
        very interesting! this seems to suggest that signal phasing and basecalling may be contributing more to the reduction in quality near the end of reads than the chemistry/imaging. if that's true then algorithms must be the way forward to 400+400 and beyond. makes me wonder if we'll ever see this chemistry on a bigger hiseq-like instrument.

        Comment

        • GenoMax
          Senior Member
          • Feb 2008
          • 7142

          #5
          Eco: I assume this run must have finished by now.

          Can we get an update?

          Comment

          • pmiguel
            Senior Member
            • Aug 2008
            • 2328

            #6
            Not sure if this would affect MiSeqs with the hardware upgrade, but there is a bug in the MCS v2.0 software that is delaying its release. I think it should have gone live Monday of this week. Word is that it will happen early next week.

            (MCS v2.0 is apparently necessary to obtain 250 cycle reads using the new v2 MiSeq chemistry reagent kits.)

            --
            Phillip

            Comment

            • Bucky
              Member
              • Feb 2010
              • 18

              #7
              Originally posted by pmiguel View Post
              Not sure if this would affect MiSeqs with the hardware upgrade, but there is a bug in the MCS v2.0 software that is delaying its release. I think it should have gone live Monday of this week. Word is that it will happen early next week.

              (MCS v2.0 is apparently necessary to obtain 250 cycle reads using the new v2 MiSeq chemistry reagent kits.)

              --
              Phillip
              yep, heard the same thing today after talking to customer support, since I wasn't able to find the software update on myillumina.

              Comment

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