I had a customer that submitted some antibody libraries which I ran on our MiSeq. The results looked good up until about 100 bases then the Q30 took a nose dive and the %A increased, suggesting that the sequence was reading into the adapter on the other side. The gel image showed a library fragment size of about 600 bp, so I had her run it on the Bioanalyzer (high sensitivity DNA). The Bioanalyzer showed a size of about 200 bp, which would be consistent with the MiSeq's poor results, but also showed a large smear at the upper marker, which doesn't show up on the gel image. Can anyone explain why the bioanayzer and agarose gel analyses would give such different results? I have attached a picture of the gel and a bioanalyzer trace of one of the samples
Thanks
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