Reducing amplification artifacts in high multiplex amplicon sequencing by using molecular barcodes.

ABSTRACT:

Background

PCR amplicon sequencing has been widely used as a targeted approach for both DNA and RNA sequence analysis. High multiplex PCR has further enabled the enrichment of hundreds of amplicons in one simple reaction. At the same time, the performance of PCR amplicon sequencing can be negatively affected by issues such as high duplicate reads, polymerase artifacts and PCR amplification bias. Recently researchers have made some good progress in addressing these shortcomings by incorporating molecular barcodes into PCR primer design. So far, most work has been demonstrated using one to a few pairs of primers, which limits the size of the region one can analyze.

Results

We developed a simple protocol, which enables the use of molecular barcodes in high multiplex PCR with hu

SUBMITTER: Peng Q 

PROVIDER: S-EPMC4528782 | biostudies-literature | 2015 Aug

REPOSITORIES: biostudies-literature