Real-time PCR quantification using a variable reaction efficiency model

Enhanced analysis of real-time PCR data by using a variable efficiency model: FPK-PCR

Current methodology in real-time Polymerase chain reaction (PCR) analysis performs well provided PCR efficiency remains constant over reactions. Yet, small changes in efficiency can lead to large quantification errors. Particularly in biological samples, the possible presence of inhibitors forms a challenge. We present a new approach to single reaction efficiency calculation, called Full Proces...

Statistical methods for efficiency adjusted real-time PCR quantification.

The statistical treatment for hypothesis testing using real-time PCR data is a challenge for quantification of gene expression. One has to consider two key factors in precise statistical analysis of real-time PCR data: a well-defined statistical model and the integration of amplification efficiency (AE) into the model. Previous publications in real-time PCR data analysis often fall short in int...

Quantification and Optimization of Candida albicans DNA in Blood Samples Using Real- Time PCR

Background: Candida albicans (C. albicans) is a major cause of candidaemia in people with impaired immunity. Blood culture is a “gold standard” for candidaemia detection but is time-consuming and relatively insensitive. We established a real-time PCR assay for C. albicans detection in blood by LightCycler PCR and melting curve analysis. Methods: Five milliliter blood samples from...

DNA quantification using EvaGreen and a real-time PCR instrument.

DNA quantiWcation is an important, frequently used technique, and inaccuracies can result in failures with ligation, restriction, polymerase chain reaction (PCR),1 ampliWed fragment length polymorphism (AFLP), Southern blotting, and other techniques. DNA is most commonly quantiWed using absorbance at 260 nm, but because of the existence of many impurities, this can be an imprecise measurement a...

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