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  • Kultamuna2
    Junior Member
    • Mar 2018
    • 5

    #1

    Sequencing process

    Hi! I am trying to find the fastest sequencing machine. How long does the sequencing take at minimum? Does the bridge amplification, sequencing, or data processing take most time? Why does the sequencing take time, if there is only 250 bases to sequence (as all reads are read simultaneously)? Thanks in advance for the comments!
  • nucacidhunter
    Jafar Jabbari
    • Jan 2013
    • 1250

    #2
    Minimum run time on Illumina systems is 4 hours on MiniSeq and MiSeq for 36 cycles. Generally, sequencing takes long time. In every cycles depending on 1, 2 or 4 colour chemistry few steps is required which includes base incorporation, imaging, debolcking and washing. YouTube search should find videos on different Illumina chemistries. One colour chemistry have slightly different workflow.

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    • snetmcom
      Senior Member
      • Oct 2008
      • 159

      #3
      Originally posted by Kultamuna2 View Post
      Hi! I am trying to find the fastest sequencing machine. How long does the sequencing take at minimum? Does the bridge amplification, sequencing, or data processing take most time? Why does the sequencing take time, if there is only 250 bases to sequence (as all reads are read simultaneously)? Thanks in advance for the comments!
      the fastest sequencing time for an average run is probably Oxford Nanopore or Ion Torrent. Both of those systems do not use optical detection or enzymatic steps to build the sequences. I suppose you could say PacBio is rather quick as well, though it does use optics.
      Last edited by snetmcom; 03-07-2018, 10:27 PM.

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      • Kultamuna2
        Junior Member
        • Mar 2018
        • 5

        #4
        Originally posted by nucacidhunter View Post
        Minimum run time on Illumina systems is 4 hours on MiniSeq and MiSeq for 36 cycles. Generally, sequencing takes long time. In every cycles depending on 1, 2 or 4 colour chemistry few steps is required which includes base incorporation, imaging, debolcking and washing. YouTube search should find videos on different Illumina chemistries. One colour chemistry have slightly different workflow.
        Thanks for the answers! Some additional questions woke in my mind when I read the answer. So at fastest sequencing takes 4 hours, does it include also the data processing (genome assembly)? Is it usually included in these given sequencing times? Is this 4 hours for a human genome?

        I have also heard that read length affects the sequencing time: shorter the read length, faster the sequencing. Why isn't then all sequencing carried out with this 36bp read length? Do we get lower quality data with lower read length?
        Last edited by Kultamuna2; 03-12-2018, 03:38 PM.

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