Question: How 2-channel sequencing chemistry (Next-Seq) distinguish "G" and 'no signal'
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gravatar for CY
22 months ago by
CY460
United States
CY460 wrote:

We found some samples enriched with "G" and we think this may caused by 2-channel sequencing chemistry can't distinguishing "G" and 'no signal'.

However, we have not have chance to sequence lot of sample using Next-Seq. Is 'no signal' or 'N' never appears in Next-Seq output? This seems to be a really serious flaw to me. Does it make sense?

qc next-seq • 681 views
ADD COMMENTlink modified 22 months ago by Devon Ryan93k • written 22 months ago by CY460
1

See for more on this issue.

ADD REPLYlink written 22 months ago by genomax76k
3
gravatar for Devon Ryan
22 months ago by
Devon Ryan93k
Freiburg, Germany
Devon Ryan93k wrote:

You will commonly see an enrichment of GGGGGGGG sequences on a NextSeq for the reason you mentioned. As an example, when you demultiplex a run you will typically find GGGGGGGG to be the "unplaced barcode" with the highest count. This is, of course, actually NNNN. Yes, this is a known limitation of NextSeq machines. For that reason I wouldn't use them for things like BS-seq, where an individual G can have a lot of meaning.

ADD COMMENTlink written 22 months ago by Devon Ryan93k
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