Biostar Beta. Not for public use.
Many 35nt reads with N bases in R1(paired-end) when sequencing with Illumina NextSeq
1
Entering edit mode
14 months ago
mgdrnl • 10
Barcelona

Dear All

We recently had a Quality Control issue when sequencing RNAseq with polyA capture in Illumina NextSeq. The reads were paired-end (2x75bp) and we found a proportion between 6%-9% of 35nt length reads with all N bases. It is also surprising that those reads all belong to the R1 reads of the paired-end library, the R2 reads look more or less ok.

enter image description here

Anybody had the same experience and know the possible technical explanation?

ADD COMMENTlink
1
Entering edit mode

What did the sequencing facility have to say about this?

To me it seems like there was some sort of failure (software/hardware/reagents) with this run. Oddly the sequencer seems to have recovered from this failure half-way during read 1 (since you said read 2 looked ok).

Depending on the answer from the sequencing facility you would be within your rights to ask that they resequence the samples (if the problem was technical and not related to your libraries).

ADD REPLYlink
0
Entering edit mode

Is there some adapter in your library ? If yes is it 35nt long ?

ADD REPLYlink

Login before adding your answer.

Similar Posts
Loading Similar Posts
Powered by the version 2.1