Evaluation of relative quantification of alternatively spliced transcripts using droplet digital PCR.

ABSTRACT:

Introduction

For the relative quantification of isoform expression, RT-qPCR has been the gold standard for over a decade. More recently, digital PCR is becoming widely implemented, as it is promised to be more accurate, sensitive and less affected by inhibitors, without the need for standard curves. In this study we evaluated RT-qPCR versus RT-droplet digital PCR (ddPCR) for the relative quantification of isoforms in controls and carriers of the splice site mutation BRCA1 c.212+3A>G, associated with increased expression of several isoforms.

Materials and methods

RNA was extracted from EBV cell lines of controls and heterozygous BRCA1 c.212+3A>G carriers. Transcript-specific plasmids were available to determine the efficiency, precision, reproducibility and accu

SUBMITTER: Van Heetvelde M 

PROVIDER: S-EPMC5634819 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature