The Renaissance of Competitive PCR as an Accurate Tool for Precise Nucleic Acid Quantification

摘要: Here, we report a detailed procedure for the exact quantification of minute amounts nucleic acids by competitive PCR. This technique entails co-amplification target DNA or cDNA in biological sample together with known quantity target-specific standard, competitor, which is added exogenously to and almost identical region be amplified. Competitive PCR offers advantage render reaction independent number amplification cycles, since any intra-assay variation has same effect on both competitor. Since final ratio between competitor products exactly reflects initial two species, being amount known, allows determination molecules target, an accuracy that still unsurpassed other alternative procedure, including real-time The protocols described this chapter cover most possible applications PCR, mRNA transcript simultaneous multiple targets.