Real-Time PCR

QPCR graph

Fluorescence-based real-time reverse transcription PCR is a widely used method for the quantification of mRNA levels in order to characterize or confirm gene expression patterns, or to compare mRNA levels of different sample populations (Bustin S.A., 2002). Real-time PCR allows for the direct detection of PCR products, combining amplification and detection in a single step. The sensitivity of this method permits the reliable detection of small amounts of initial template, while delivering a linear range of up to ten orders of magnitude in copy number. A typical amplification plot of fluorescent intensities over 40 cycles is shown here.

The Bauer Core provides access to several real time PCR instruments that support all of the popular real-time chemistries. Our staff helps users with experimental design and provides training on the instruments.

For more information about real time PCR at the Bauer Core, please contact Claire Reardon (claire@cgr.harvard.edu).

Reference:

Bustin SA. Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. Journal of Molecular Endocrinology (2002) 29: 23–39