If the linear amplification profile of the library is significantly below fluorescent standard 1 at the end of qPCR cycling, then it is unlikely that there will be sufficient library material to sequence after PCR purification.
If the linear amplification profile of the library is significantly above fluorescent standard 3 at the end of the qPCR cycling, then the library has been over-amplified. This may lead to amplification bias, higher error rates, and/or the presence of chimeric PCR products.
2017-12-21