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Genome biology DOI:10.1186/gb-2011-12-2-r18

Analyzing and minimizing PCR amplification bias in Illumina sequencing libraries.

Publication TypeJournal Article
Year of Publication2012
AuthorsAird, D, Ross, MG, Chen, WS, Danielsson, M, Fennell, T, Russ, C, Jaffe, DB, Nusbaum, C, Gnirke, A
JournalGenome biology
Volume12
Issue2
PagesR18
Date Published2012/01/09
ISSN1465-6906
Abstract

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.

URLhttp://genomebiology.com/content/12/2/R18
DOI10.1186/gb-2011-12-2-r18
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/21338519?dopt=Abstract