A robust internal control for high-precision DNA methylation analyses by droplet digital PCR.

ABSTRACT:

Background

Droplet digital PCR (ddPCR) allows absolute quantification of nucleic acids and has potential for improved non-invasive detection of DNA methylation. For increased precision of the methylation analysis, we aimed to develop a robust internal control for use in methylation-specific ddPCR.

Methods

Two control design approaches were tested: (a) targeting a genomic region shared across members of a gene family and (b) combining multiple assays targeting different pericentromeric loci on different chromosomes. Through analyses of 34 colorectal cancer cell lines, the performance of the control assay candidates was optimized and evaluated, both individually and in various combinations, using the QX200™ droplet digital PCR platform (Bio-Rad). The best-performing control wa

SUBMITTER: Pharo HD 

PROVIDER: S-EPMC5822558 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature