Analyzing and minimizing PCR amplification bias in Illumina sequencing libraries View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2011-02

AUTHORS

Daniel Aird, Michael G Ross, Wei-Sheng Chen, Maxwell Danielsson, Timothy Fennell, Carsten Russ, David B Jaffe, Chad Nusbaum, Andreas Gnirke

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. More... »

PAGES

r18

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/gb-2011-12-2-r18

DOI

http://dx.doi.org/10.1186/gb-2011-12-2-r18

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1000268052

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/21338519


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