Selective control of primer usage in multiplex one-step reverse transcription PCR.

ABSTRACT:

Background

Multiplex RT-PCR is a valuable technique used for pathogen identification, disease detection and relative quantification of gene expression. The simplification of this protocol into a one-step procedure saves time and reagents. However, intensive PCR optimization is often required to overcome competing undesired PCR primer extension during the RT step.

Results

Herein, we report multiplex one-step RT-PCR experiments in which the PCR primers contain thermolabile phosphotriester modification groups. The presence of these groups minimizes PCR primer extension during the RT step and allows for control of PCR primer extension until the more stringent, elevated temperatures of PCR are reached. Results reveal that the use of primers whose extension can be controlled in a

SUBMITTER: Hidalgo Ashrafi E 

PROVIDER: S-EPMC2811113 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature