|Publication Type||Journal Article|
|Year of Publication||2012|
|Authors||Aird, D, Ross, MG, Chen, WS, Danielsson, M, Fennell, T, Russ, C, Jaffe, DB, Nusbaum, C, Gnirke, A|
Despite the ever-increasing output of Illumina sequencing data, loci with extreme base compositions are often under-represented or absent. To evaluate sources of base-composition bias, we traced genomic sequences ranging from 6% to 90% GC through the process by quantitative PCR. We identified PCR during library preparation as a principal source of bias and optimized the conditions. Our improved protocol significantly reduces amplification bias and minimizes the previously severe effects of PCR instrument and temperature ramp rate.