Influence of PCR Reagents on DNA Polymerase Extension Rates Measured on Real-Time PCR Instruments

Influence of PCR reagents on DNA polymerase extension rates measured on real-time PCR instruments.

BACKGROUND Radioactive DNA polymerase activity methods are cumbersome and do not provide initial extension rates. A simple extension rate assay would enable study of basic assumptions about PCR and define the limits of rapid PCR. METHODS A continuous assay that monitors DNA polymerase extension using noncovalent DNA dyes on common real-time PCR instruments was developed. Extension rates were ...

Influence of reagents formulation on real-time PCR parameters.

Real-time polymerase chain reaction (PCR) techniques are increasingly used to quantify target sequences for diagnostic and research purposes. Due to its 'quantitative' character, it is very important to determine the variability of this technique correlating with several experimental conditions. The objective of this study was to analyse the effect of manufacturing lots of PCR reagents on two m...

Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Fatemeh Mehrpooyan*

Real-time PCR detection of parapoxvirus DNA.

BACKGROUND Detection of parapoxviruses is important in various animals as well as in humans as zoonotic infections. Reliable detection of parapoxviruses is fundamental for the exclusion of other rash-causing illnesses, for both veterinarians and medical practitioners. To date, however, no real-time PCR assay for the detection of parapoxviruses has been reported. METHODS A minor groove binder-...

Influence of DNA target melting behavior on real-time PCR quantification.

BACKGROUND Quantitative real-time PCR is increasingly used to quantify copy numbers of nucleic acids for clinical applications. We observed that the measurements of allele imbalances of the tumor suppressor gene p16 and the oncogene ErbB-2 yielded results with variable precision under certain experimental conditions. METHODS We used the LightCycler(TM) real-time PCR system to quantify differe...