Scientific Publications

Analyzing and minimizing PCR amplification bias in Illumina sequencing libraries.

Publication TypeJournal Article
AuthorsAird, D., Ross MG, Chen WS, Danielsson M., Fennell T., Russ C., Jaffe DB, Nusbaum C., and Gnirke A.
AbstractDespite 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.
Year of Publication2012
JournalGenome biology
Date Published (YYYY/MM/DD)2012/01/09
ISSN Number1465-6906