Research Reports BioTechniques研究报告生物技术.pdfVIP

Research Reports Reproducibility in the Quantification of mRNA Levels by RT-PCR-ELISA and RT Competitive- PCR-ELISA BioTechniques 24:652-658 (April 1998) L.L. Hall, G.R. Bicknell, L. INTRODUCTION be appropriate where there is a limited Primrose, J.H. Pringle, J.A. amount of mRNA from microdissected Shaw and P.N. Furness Quantification of levels of mRNA is tissue samples. University of Leicester, important in the study of gene expres- RT-competitive PCR (RT-cPCR) re- sion. Reverse transcription polymerase quires the addition of an internal com- Leicester, England, UK chain reaction (RT-PCR) is widely used petitive DNA standard to the PCR after in the investigation of low-copy-num- completion of the RT step (8). This ber mRNAs and all mRNAs from small method permits the PCR amplification samples (3,5,7,9). The sensitivity of and analysis of multiple genes from a RT-PCR has enabled the analysis of mi- single RT reaction, and it overcomes ABSTRACT crodissected human tissue samples, any problems with the stability of an providing some degree of localization. RNA competitor. However, it assumes The use of reverse transcription (RT) What remains to be investigated is the constant efficiency of the RT reaction. PCR for