SmaRT-PCR: a novel application of competitive PCR for mRNA quantitation

Abstract

The polymerase chain reaction (PCR) was first described in 1985 (Ref. [1]). Since then, it has become the ‘gold standard’ for amplifying DNA and reverse-transcribed RNA. Unfortunately, the greatest advantage of PCR (its sensitivity) easily becomes its biggest disadvantage, as the smallest changes in reaction conditions from tube to tube result in large differences in the outcome.

During the past few years, there has been increasing interest in semiquantitative and quantitative PCR (Ref. [2]). Several protocols for how to generate and use internally deleted standards have been published (Ref. 3, 4, 5). The aim of this study was to measure gene expression using the high specificity and sensitivity of competitive PCR techniques but at a lower cost.