Brief Introduction to PCR-Pharmaceutical Biotechnology-Unit 2- B. Pharmacy 6 Sem-Lect.7

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#Brief_Introduction_To_PCR #Pharmaceutical_Biotechnology
The polymerase chain reaction PCR is a laboratory (in vitro) technique for generating large quantities of a specified DNA. Obviously PCR is a cell free amplification technique for synthesizing multiple identical copies (billions) of any DNA of interest. It was developed in 1984 by karry mullis (Nobel laureate 1993).
Principle of PCR:
The double stranded DNA of interest is denatured to separate into two individual strands. Each strand is then allowed to hybridize with the primer (renaturation). The primer template duplex is used for DNA synthesis (the enzyme DNA polymerase).

Technique of PCR-the essential requirements for PCR are listed below-
A target DNA 100 to 35000 base pair in length
Two primers synthetic oligonucleotides of 17-30 nucleotide lengths that are complementary to regions flanking the target DNA.
Four types of deoxyribonucleotides (dATP, dCTP, dGTP, dTTP)
A DNA polymer can withstand at a temperature up to 95° C that is thermostable.


Each cycle has three stages-
1) Denaturation
2) Renaturation or annealing
3) Synthesis

Key factors for optimal PCR
1) Primers
2) DNA polymerase
3) Target DNA
4) Promoters and inhibitors

Application of PCR







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