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  • Poor loading on Ion Proton

    We've suddenly found loading to be really poor on our Ion proton. Previously it was around 85-90%, with 80-90+ %, with 70-90M reads. Suddenly it is as low as 42% with only around 20M reads.

    Has anyone got ideas as to why this might be? Is it likely to be chip-prep related? Can the library affect it?

    A

  • #2
    All of the above. It this a library you have sequenced well before, or is it a completely new experiment.

    Comment


    • #3
      Have you done by any chance a Qbit on your unenriched template?
      Low number for both dyes, could mean that you have lost some beads in the process prior to running the ES.

      Remito

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      • #4
        any new update? we have this problem recently.

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        • #5
          Poor loading could be caused by,

          -Removing too much sample during the enrichment process by accidentally pippetting out more than the suggested amount
          -Not spinning the enriched sample and accidentally removing the beads during the last step of enrichment
          -Not enough beads added prior to emulsion PCR

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