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Old 08-08-2014, 09:20 AM   #1
ScottishSequencer
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Default 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
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Old 08-10-2014, 03:12 PM   #2
snetmcom
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All of the above. It this a library you have sequenced well before, or is it a completely new experiment.
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Old 08-15-2014, 01:10 PM   #3
RemitoAmigo
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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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Old 08-24-2014, 08:34 PM   #4
feng
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any new update? we have this problem recently.
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Old 10-08-2014, 04:50 PM   #5
NextGenUser
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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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