Hi everybody,
I tried for the first time the Clontech Stranded mRNA-Seq Kit for Illumina to generate libraries. My starting material (1 ng total RNA isolated from human serum) was ok. However, after library preparation I ended up with a weird looking electropherogram on the Bioanalyzer. My first thought was that this might be due to overamplification, so I reduced the final amplification cycles (12-14-16). Unfortunately, this didn’t solve the problem.
Do anyone has an idea what’s with these libraries with the expected peak at 300 and the additional peaks at ~400 and 600?
PS. Fragmentation time was 2 minutes (I also tried 4 minutes)
Thank you.
I tried for the first time the Clontech Stranded mRNA-Seq Kit for Illumina to generate libraries. My starting material (1 ng total RNA isolated from human serum) was ok. However, after library preparation I ended up with a weird looking electropherogram on the Bioanalyzer. My first thought was that this might be due to overamplification, so I reduced the final amplification cycles (12-14-16). Unfortunately, this didn’t solve the problem.
Do anyone has an idea what’s with these libraries with the expected peak at 300 and the additional peaks at ~400 and 600?
PS. Fragmentation time was 2 minutes (I also tried 4 minutes)
Thank you.
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