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Old 10-16-2019, 06:20 AM   #1
acockburn
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Location: Pa

Join Date: Aug 2013
Posts: 10
Question NextSeq underclustering

Has anyone had issues with underclustering on the Illumina NextSeq 500/550? We have been having issues all summer. Even increasing input 3 times our normal has not caused overclustering. We usually clustered around 200 K/mm2, now we typically get 100-140 with 3 times the amount. It is not overclustered as we still get good PF and Q30 scores.

We have checked the qPCR, the quant kits, all the reagents, seems the only variable left is the NextSeq.

We think it may have started with a change to the v2.5 Flowcells, but are uncertain.

Anybody experiencing this? Anybody have any suggestions?

Thanks.
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