Single-tube, highly parallel mutation enrichment in cancer gene panels by use of temperature-tolerant COLD-PCR.

ABSTRACT:

Background

Multiplexed detection of low-level mutations presents a technical challenge for many technologies, including cancer gene panels used for targeted-resequencing. Analysis of mutations below approximately 2%-5% abundance in tumors with heterogeneity, samples with stromal contamination, or biofluids is problematic owing to increased noise from sequencing errors. Technologies that reduce noise via deep sequencing unavoidably reduce throughput and increase cost. Here we provide proof of principle that coamplification at lower denaturation temperature (COLD)-PCR technology enables multiplex low-level mutation detection in cancer gene panels while retaining throughput.

Methods

We have developed a multiplex temperature-tolerant COLD-PCR (fast-TT-COLD-PCR) approach that use

SUBMITTER: Castellanos-Rizaldos E 

PROVIDER: S-EPMC4281501 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature