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Polymerase Chain Reaction

  • Alangar Ishwara Bhat
  • Govind Pratap Rao
Protocol
  • 44 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Polymerase chain reaction (PCR) is the most widely used nucleo-based method for the detection of plant viruses. It is a primer-mediated in vitro reaction involving amplification of target nucleic acid sequences. A standard PCR is a three-step procedure: (1) denaturation at a high temperature (90–95 °C), (2) annealing of target specific primers (45–60 °C) and (3) primer extension by a thermostable DNA polymerase at 72 °C. Different variants of PCR have been developed ranging from conventional PCR to real-time PCR to improve the sensitivity and specificity for the detection of plant viruses. Some virus detection methods are: DNA-based PCR, nested PCR (nPCR), immunocapture PCR (IC-PCR), multiplex PCR (M-PCR), real-time PCR and DNA finger printing. Others are RNA-based reverse transcription (RT) PCR, real-time RT-PCR, IC-RT-PCR and AmpliDet RNA. All these methods enable a rapid and accurate detection and quantification of plant viruses. Protocols of PCR assays for the detection of plant viruses are discussed in this chapter.

Key words

PCR Immunocapture PCR Nested PCR RT-PCR IC-RT-PCR 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Alangar Ishwara Bhat
    • 1
  • Govind Pratap Rao
    • 2
  1. 1.Division of Crop ProtectionICAR-Indian Institute of Spices ResearchKozhikodeIndia
  2. 2.Division of Plant PathologyICAR-Indian Agricultural Research InstituteNew DelhiIndia

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