Contamination aside, have any of you looked at the effect of resampling bias from doing so much PCR on a sample? I have a customer looking to do this approach coming from RNA. This means 4 rounds of PCR, 1 RT reaction, 1 target specific nest, then the Illumina 2 step. I plan on titrating yield on a Lonza gel to optimize PCR cycles, but still sounds like a lot of resampling.
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