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  • Kultamuna2
    Junior Member
    • Mar 2018
    • 5

    #1

    Illuminas operating system

    Hi! Can anyone tell me, why Illumina's sequencing systems have two lasers (or in NovaSeq, four lasers)? I am a bit confused how the system works. Do those lasers change constantly wavelengths to identify all the four bases, or do they stay in a specific wavelength? Thanks already in advance!
  • kc3228
    Junior Member
    • May 2017
    • 6

    #2
    The two lasers are red and green. Red is C, green is T. A is labeled with both red and green, so shows up in both. G is dark (unlabeled), so a lack of signal is interpreted as G.

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    • GW_OK
      Senior Member
      • Sep 2009
      • 411

      #3
      The lasers excite at fixed wavelengths. The fluorophores emit at different respective wavelengths. The example given by kc3228 is for the dual color chemistries of the NextSeq and NovaSeq. The MiniSeq, Miseq, and Hiseq systems use a 4 color chemistry.

      Comment

      • misterc
        Member
        • Jan 2016
        • 46

        #4
        MiniSeq is essentially a single camera NextSeq, so 2-color SBS there (not 4).

        Comment

        • GW_OK
          Senior Member
          • Sep 2009
          • 411

          #5
          Good to know.

          I suppose the iSeq is single color, since it's got the CMOS in the flowcell.

          Comment

          • GenoMax
            Senior Member
            • Feb 2008
            • 7142

            #6
            iSeq is one-dye, two-chemistry and two-images per sequencing cycle. See page 3.

            Comment

            • kmcarr
              Senior Member
              • May 2008
              • 1181

              #7
              Here's another doc from Illumina which also describes the CMOS flow cell and compares the various detection chemistries (1, 2, 4 color) used by Illumina.

              Comment

              • GenoMax
                Senior Member
                • Feb 2008
                • 7142

                #8
                Nice find @kmcarr. Thanks for posting.

                Comment

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