qScript XLT cDNA SuperMix: overcoming the common pitfalls of cDNA synthesis as applied to relative quantification and RT-qPCR

David Mark Schuster, Yun Feng Quanta BioSciences, United States of America

Abstract
Despite the introduction of new methods for gene expression quantification, reverse transcription quantitative PCR (RT-qPCR) remains the dominant method for measuring changes in gene expression, providing high accuracy and precision with broad dynamic range. At the heart of this technique is the reverse transcription of RNA into complementary first-strand DNA product. The ready availability of reliable and easy to use kits has made this critical step of the process routine. However, complacency and the lack of attention to this critical portion of the process can be at the peril of robust and reliable data and introduce potential bias that can misleadingly be attributed to differential gene expression.
This talk will examine a variety of factors that influence RT-qPCR and present an advanced first-strand synthesis system that offers significant improvement in the sensitivity and accuracy of RT-qPCR for routine applications as well as the demanding needs of reverse transcription from limited template amounts such as the study of gene expression in single cells.