Journal
GENOME BIOLOGY
Volume 12, Issue 2, Pages -Publisher
BIOMED CENTRAL LTD
DOI: 10.1186/gb-2011-12-2-r18
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Funding
- National Institute of Allergy and Infectious Diseases [HHSN27220090018C]
- National Human Genome Research Institute [HG03067-05]
- NATIONAL HUMAN GENOME RESEARCH INSTITUTE [U54HG003067, R01HG003474] Funding Source: NIH RePORTER
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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.
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