Preventing RNA degradation is difficult, but newer systems for protecting this molecule may lead the way to better gene expression analysis.

摘要

RNA integrity is crucial for obtaining reliable and meaningful gene expression data. Using highly-purified, intact RNA samples is a key component for the successful application of modern molecular biology and diagnostic techniques such as quantitative real-time PCR (qRT-PCR) or microarray analysis. qRT-PCR realizes the inherent quantitative potential of PCR, making it a quantitative, as well as qualitative, assay that is the method of choice for the detection of mRNA. While qRT-PCR has been transformed into a mainstream research tool, there are limitations to the technology. A lack of standardization and normalization can make it difficult to draw definitive conclusions about the biological validity of quantitative data. The intimate association between quantification and amplification efficiency during the reverse transcription step highlights the critical importance of using mRNA that is of high quality and integrity as the starting point for the reaction.