< Back to Publications
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|
|Date Published (YYYY/MM/DD)||2012/01/09|