Hello,
We are experiencing an issue with the TruSeq® Stranded mRNA HT kit. We prepared three 96-well plates of RNAseq libraries in our EpMotion robots. The libraries are very consistent and have approximately the same concentration. We first sequenced one pool of 96 libraries on 8 lanes with single-reads 100nt, v4 chemistry. We saw that the libraries coming from column 3, which all have adaptor D703, had low number of reads and the reads from these libraries had low quality scores, whereas all the other libraries had great consistency in number of reads and quality.
The same thing happened with the other pools, always the libraries coming from this column did not perform well. Libraries were done in different robots, different months and different kit lot numbers.
We called Illumina and they asked us to sequence the same pools with the V3 chemistry and they provided the kits. And voila! All libraries sequenced beautifully.
There has been no resolution on this yet as we are only the 3rd lab to report this problem.
Has anyone else seen this issue?
Thanks,
Alvaro
We are experiencing an issue with the TruSeq® Stranded mRNA HT kit. We prepared three 96-well plates of RNAseq libraries in our EpMotion robots. The libraries are very consistent and have approximately the same concentration. We first sequenced one pool of 96 libraries on 8 lanes with single-reads 100nt, v4 chemistry. We saw that the libraries coming from column 3, which all have adaptor D703, had low number of reads and the reads from these libraries had low quality scores, whereas all the other libraries had great consistency in number of reads and quality.
The same thing happened with the other pools, always the libraries coming from this column did not perform well. Libraries were done in different robots, different months and different kit lot numbers.
We called Illumina and they asked us to sequence the same pools with the V3 chemistry and they provided the kits. And voila! All libraries sequenced beautifully.
There has been no resolution on this yet as we are only the 3rd lab to report this problem.
Has anyone else seen this issue?
Thanks,
Alvaro
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