Journal article

A one-step quantitative reverse transcription polymerase chain reaction procedure

KL Kelleher, KJ Leck, IA Hendry, KI Matthaei

BRAIN RESEARCH PROTOCOLS | ELSEVIER SCIENCE BV | Published : 2001

Abstract

Our laboratory has developed a one-step quantitative reverse transcription polymerase chain reaction (RT-PCR) procedure in which the reverse transcriptase enzyme and Taq DNA polymerase are combined in the one tube and a single, non-interrupted, thermal cycling program is performed. In the past, RT-PCR has been carried out with two separate steps: (1) reverse transcription of RNA to generate a cDNA pool and (2) polymerase chain reaction amplification of the cDNA. The two-step method can affect the accuracy of the procedure as the total number of manipulations is greater, thereby allowing a greater chance for pipetting errors. Quantitation by our method is achieved in a single reaction by the ..

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