Professional Documents
Culture Documents
Abstract
nYQp/IlQrHD3i3D0OdRyi7TvSFl4Cf3VC4/OAVpDDa8K2+Ya6H515kE= on 02/26/2024
DNA extraction and polymerase chain reaction (PCR) are the basic techniques employed in the molecular laboratory. This short overview
covers various physical and chemical methods used for DNA extraction so as to obtain a good‑quality DNA in sufficient quantity. DNA can
be amplified with the help of PCR. The basic principle and different variants of PCR are discussed.
Access this article online This is an open access journal, and articles are distributed under the terms of the Creative
Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to
Quick Response Code: remix, tweak, and build upon the work non-commercially, as long as appropriate credit
Website: is given and the new creations are licensed under the identical terms.
www.jcytol.org
For reprints contact: reprints@medknow.com
DOI:
10.4103/JOC.JOC_110_18 How to cite this article: Gupta N. DNA extraction and polymerase chain
reaction. J Cytol 2019;36:116-7.
116 © 2019 Journal of Cytology | Indian Academy of Cytologists | Published by Wolters Kluwer - Medknow
Gupta: DNA extraction polymerase chain reaction
Polymerase Chain Reaction Hot‑start PCR: The main advantage of hot‑start PCR is
to decrease nonspecific amplification of DNA at lower
Polymerase chain reaction (PCR) is a robust technique to
temperature steps of PCR. Reaction components are manually
selectively amplify a specific segment of DNA in vitro.[1] PCR is
heated before adding Taq polymerase to the DNA‑melting
performed on thermocycler and it involves three main steps: (1)
temperature (i.e. 95°C).[4]
denaturation of dsDNA template at 92–95°C, (2) annealing of
primers at 50–70°C, and (3) extension of dsDNA molecules Touchdown PCR: Annealing temperature during the first
Downloaded from http://journals.lww.com/jocy by BhDMf5ePHKav1zEoum1tQfN4a+kJLhEZgbsIHo4XMi0hCywCX1AW
at approx. 72°C. These steps are repeated for 30–40 cycles. two cycles of amplification is set at approximately 3–10°C
above estimated Tm and the temperature is slowly decreased
Various chemical components of PCR include MgCl 2,
in the subsequent cycles. Higher annealing temperature in
buffer (pH: 8.3–8.8), Deoxynucleoside triphosphates
two initial cycles leads to more specificity for primer binding,
(dNTPs), PCR primers, target DNA, and thermostable DNA
nYQp/IlQrHD3i3D0OdRyi7TvSFl4Cf3VC4/OAVpDDa8K2+Ya6H515kE= on 02/26/2024