Reverse Transcriptase (RT) Inhibition of PCR at Low Concentrations of Template and Its Implications for Quantitative RT-PCR

Reverse transcriptase (RT) inhibition of PCR at low concentrations of template and its implications for quantitative RT-PCR.

Numerous instances of reverse transcriptase (RT) inhibition of the PCR were observed while developing nonquantitative uncoupled RT-PCR techniques for detecting nitrogenase and ammonia monooxygenase gene expression in situ. The inhibitory effect of RT on the PCR was removed with increasing template concentrations beyond 10(5) to 10(6) copies. Including T4 gene 32 protein during the reverse trans...

Reverse transcriptase-polymerase chain reaction (rt-PCR).

Reverse transcriptase polymerase chain reaction (RT-PCR) has become a well-established and powerful molecular technique for studying ribonucleic acids. It is used in medical diagnostics for detecting viral RNA, in hematology and oncology for detecting chimeric transcripts of rearranged genes (1), and in the broad area of research applications in gene expression studies. Since its introduction (...

Quantitative PCR and RT-PCR in virology.

I n the last few years, molecular hybridization methods have been used extensively in basic and applied virology because of their technical flexibility and high specificity. Using these techniques, the detection of DNA and RNA viruses directly from clinical specimens, the analysis of the specific transcriptional activity of viral genes in vitro and in vivo, and the study of virus-host relations...

Guidelines for quantitative rt-PCR.

Something I notice often in supervising new graduate students is their frustration with the irreproducibility of their biological data. For example, I am asked frequently ‘‘how many biological replicates do I need for qRT-PCR’’? The answer, of course, lies first with the students themselves. They need as many replicates as will persuade them of the validity of their observations. However, for m...

One-Step Reverse Transcription PCR (RT-PCR). Fatemeh Mehrpooyan*