Quantitative PCR for cAMP RI Alpha mRNA: Use of Site-Directed Mutation and PCR Mimics

Precise and accurate determination of mRNA expression levels in tissues and model systems is a central methodology in a wide range of research applications. Expression of many genes is currently assessed by northern blotting, RNAse protection assays, Serial Analysis of Gene Expression (SAGE), and many other techniques; however, for single transcripts, especially where tissue is limited or abundance is low, quantitative polymerase chain reaction (PCR) is the method of choice. However, where quantitative PCR is to be used, the reproducibility, accuracy, and detection limits of the technique must be clearly defined.