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Polymerase Chain Reaction: A Robust Technology for Crop Improvement
Vivekanand Tiwari*, Kiran Patel, Viral B. Mandaliya, and Sujit Kumar Bishi
Directorate of Groundnut Research, PB-05, Ivnagar Road, Junagadh-362 001, Gujarat
*Email of corresponding author: vivekcas805@gmail.com
Introduction
In 1983, Karry Mullis and co-workers have developed PCR technique to replicate fragments
of DNA by semi-conservative method but under in-vitro condition. The introduction of
thermostable Taq DNA polymerase from Thermus aquaticus has facilitated automation of
this technology (Saiki et al., 1988). The technique was patented by Kary Mullis at Cetus
Corporation, where he invented. The use of this invention for crop improvement was widely
accepted in the areas of research like detection of plant pathogens, development of genetic or
physical map of crop genome, cloning and characterization of existing or new trait responsive
genes and their expression pattern analysis, genetic diversity analysis and marker assisted
breeding for development of new crop varieties. Although, PCR has become a routine tool in
molecular biology research for crop improvement studies, but still there are few important
points should be taken care while performing a PCR. Different modifications in the
components and reaction conditions have been adopted according to the applications or aim
of the experiments for successful PCR reaction. This articles focus on the basic principle of
PCR, what are the precautions should be taken care and application of PCR for crop
improvements.
Principle of PCR
The principle of a PCR reaction can be generalized as the, in-vitro replication of small
fragment of template DNA upto a desired size (< 10 kb). The DNA product formed after PCR
are known as amplicons. Amplicons are increased in number exponentially after completion
of each cycle. It includes separation of two opposite DNA strand by heat denaturation instead
of enzymatic unwinding occurs in cells. A pair of small single stranded DNA fragments
Popular Kheti
Volume -2, Issue-3 (July-September), 2014
Available online at www.popularkheti.info
© 2014 popularkheti.info
ISSN: 2321-0001
Polymerase Chain Reaction (PCR) is widely used molecular biology research tools
discovered by Kary Mullis in 1983. In this technology, with use of thermostable Taq
DNA polymerase a fragment of DNA was transcribed in-vitro. The amplification
was carried out using small fragment of DNA complementary to both ends of the
DNA to be amplified and called as primer. Primers were designed carefully to avoid
non-specific amplification. This technology widely used in crop improvement
programs like development of genetic or physical map of crop genome, cloning and
characterization of existing or new trait responsive genes and their expression
pattern analysis, genetic diversity analysis and marker assisted breeding for
development of new crop varieties.
Popular Kheti ISSN: 2321-0001 225