Dear all,
I need an advice regarding latest NextSeq 500 sequencing run. Given - NextSeq 500, HighOutput 75 cycles sequencing kit v2, highly multiplexes (192 libraries) sequencing pool of genomic DNA library samples prepared with in-house tagmentation procotol and seven additional Cut&Tag libraries. All libraries have 8bp i5 and i7 barcodes. Bioanalyzier profiles and library concentrations looks fine as usual. The DNA libraries are routine for our lab and never caused such kind of troubles before. Cut&Tag libraries are rather new test samples. I do not know why they would affect the sequencing, but they might be considered as an unknown variable in the equation.
The library pools were diluted to 2nM final concentration and denatured according standard Illumina instructions. The cluster density is about 200, which is quite typical for this machine and our sample preparation, however only 1.91% of clusters passed filter per lane giving 0.92Gb yeild. What are the first things to check? Thanks a lot in advance.
PS One of the components of the kit expired about two months ago, so I can't really contact Illumina, but this was never an issue in the past. Most sequencing kits works well even years after the expiration date.
I need an advice regarding latest NextSeq 500 sequencing run. Given - NextSeq 500, HighOutput 75 cycles sequencing kit v2, highly multiplexes (192 libraries) sequencing pool of genomic DNA library samples prepared with in-house tagmentation procotol and seven additional Cut&Tag libraries. All libraries have 8bp i5 and i7 barcodes. Bioanalyzier profiles and library concentrations looks fine as usual. The DNA libraries are routine for our lab and never caused such kind of troubles before. Cut&Tag libraries are rather new test samples. I do not know why they would affect the sequencing, but they might be considered as an unknown variable in the equation.
The library pools were diluted to 2nM final concentration and denatured according standard Illumina instructions. The cluster density is about 200, which is quite typical for this machine and our sample preparation, however only 1.91% of clusters passed filter per lane giving 0.92Gb yeild. What are the first things to check? Thanks a lot in advance.
PS One of the components of the kit expired about two months ago, so I can't really contact Illumina, but this was never an issue in the past. Most sequencing kits works well even years after the expiration date.
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