Humic substances cause fluorescence inhibition in real-time polymerase chain reaction

Taq polymerase reverses inhibition of quantitative real time polymerase chain reaction by humic acid.

AIM To investigate the dose-response effect of humic acid (HA) on the quantitative real time polymerase chain reaction (QRT-PCR) inhibition and the efficiency of Taq polymerase increment in preventing inhibition by HA in DNA extracted from ancient bones. METHODS DNA was isolated from bone samples and DNA quantification was conducted with the real-time 5' exonuclease detection assay (TaqMan), ...

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

Real-time polymerase chain reaction.

Real-time PCR is the state-of-the-art technique to quantify nucleic acids for mutation detection, genotyping and chimerism analysis. Since its development in the 1990s, many different assay formats have been developed and the number of real-time PCR machines of different design is continuously increasing. This review provides a survey of the instruments and assay formats available and discusses...

Detection of Nocardia Asteroides Complex in Clinical Isolates by Real-Time Polymerase Chain Reaction

Background and Aims: Nocardia asteroides complex is the most common cause of infectious diseases due to nocardiosis. Interspecies differentiation of Nocardia genera is essential for prognosis and timely proper treatment, as well as for epidemiological studies. Since each genus has its own antibiotic resistance, precise careful diagnosis is of prime importance. As compared to biochemical and phe...

The real-time polymerase chain reaction.

The scientific, medical, and diagnostic communities have been presented the most powerful tool for quantitative nucleic acids analysis: real-time PCR [Bustin, S.A., 2004. A-Z of Quantitative PCR. IUL Press, San Diego, CA]. This new technique is a refinement of the original Polymerase Chain Reaction (PCR) developed by Kary Mullis and coworkers in the mid 80:ies [Saiki, R.K., et al., 1985. Enzyma...