PCR ( Polymerase chain Reaction)



The polymerase chain reaction (PCR) is a scientific technique in molecular biology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. Developed in 1983 by Kary Mullis, PCR is now a common and often indispensable technique used in medical and biological research labs for a variety of applications. These include DNA cloning for sequencing, DNA-based phylogeny, or functional analysis of genes; the diagnosis of hereditary diseases; the identification of genetic fingerprints (used in forensic sciences and paternity testing); and the detection and diagnosis of infectious diseases. In 1993, Mullis was awarded the Nobel Prize in Chemistry for his work on PCR.


The method relies on thermal cycling, consisting of cycles of repeated heating and cooling of the reaction for DNA melting and enzymatic replication of the DNA. Primers (short DNA fragments) containing sequences complementary to the target region along with a DNA polymerase (after which the method is named) are key components to enable selective and repeated amplification. As PCR progresses, the DNA generated is itself used as a template for replication, setting in motion a chain reaction in which the DNA template is exponentially amplified.


PCR CONDITIONS:- 


PCR conditions may be vary according to your Experiments


Initial denaturation 94°C for 5 minutes 


Denaturation 94°C for 30 seconds 


Annealing 60°C for 40 seconds 


Extension 72°C for 30 seconds 


Final extension 72°C for 4 minutes 


Final hold 4°C.

Editor: Ankita Added on: 2021-02-26 13:40:52 Total View:334







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