1] Department of Biology, University of North Carolina, Chapel Hill, North Carolina, USA. [2] Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, North Carolina, USA. [3].
Nat Methods. 2013 Oct;10(10):999-1002. doi: 10.1038/nmeth.2634. Epub 2013 Sep 1.
We describe improvements for sequencing 16S ribosomal RNA (rRNA) amplicons, a cornerstone technique in metagenomics. Through unique tagging of template molecules before PCR, amplicon sequences can be mapped to their original templates to correct amplification bias and sequencing error with software we provide. PCR clamps block amplification of contaminating sequences from a eukaryotic host, thereby substantially enriching microbial sequences without introducing bias.
我们描述了对 16S 核糖体 RNA（rRNA）扩增子测序的改进，这是宏基因组学的一项基石技术。通过在 PCR 之前对模板分子进行独特标记，我们提供的软件可以将扩增子序列映射回其原始模板，以纠正扩增偏倚和测序错误。PCR 夹阻断了真核宿主中污染序列的扩增，从而在不引入偏倚的情况下大大富集微生物序列。
