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Analyzing and minimizing PCR amplification bias in Illumina sequencing libraries.
| Publication Type | Journal Article |
| Authors | Aird, D., Ross MG, Chen WS, Danielsson M., Fennell T., Russ C., Jaffe DB, Nusbaum C., and Gnirke A. |
| 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. |
| Year of Publication | 2012 |
| Journal | Genome biology |
| Volume | 12 |
| Issue | 2 |
| Pages | R18 |
| Date Published (YYYY/MM/DD) | 2012/01/09 |
| ISSN Number | 1465-6906 |
| DOI | 10.1186/gb-2011-12-2-r18 |
| PubMed | http://www.ncbi.nlm.nih.gov/pubmed/21338519?dopt=Abstract |
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