Poor loading on Ion Proton

[ScottishSequencer]

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

[snetmcom]

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

[RemitoAmigo]

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

[feng]

any new update? we have this problem recently.

[NextGenUser]

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