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General Principles of Real-Time PCR: A Technology for Quantitative Detection of Phytopathogens

Seyed Mahyar Mirmajlessi 1, Evelin Loit 1, Marika Mänd 2
1. Department of Field Crops and Grassland Husbandry, Estonian University of Life Sciences, Tartu, Estonia
2. Department of Plant Protection, Estonian University of Life Sciences, Tartu, Estonia
Abstract—Real-time PCR provides a rapid and specific method for detection of plant pathogens in various environmental samples. The technique incorporates traditional PCR efficiency with the production of a specific fluorescent signal, providing quantification of specific targets. There are four main chemistries, which can be clustered into the non-specific DNA-binding fluorophores and the specific fluorophore-labelled oligonucleotide probes. In the present review, we describe the general background of major real-time chemistries as well as some considerations for assay development. Basically, Scorpion and SYBR green have the most and least sensitivities between available chemistries, respectively. However, with accurate optimization, real-time PCR can provide reliable and high throughput detection of target DNA in various environments.

Index Terms—alternative chemistries, plant pathology, real-time PCR

Cite: Seyed Mahyar Mirmajlessi, Evelin Loit, and Marika Mänd, "General Principles of Real-Time PCR: A Technology for Quantitative Detection of Phytopathogens," Journal of Medical and Bioengineering, Vol. 5, No. 1, pp. 49-52, February 2016. Doi: 10.12720/jomb.5.1.49-52
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