Polymerase Chain Reaction (PCR) | OnlineBiotechNotes

Polymerase Chain Reaction (PCR)

Polymerase chain reaction by OnlineBiotechNotes

• PCR is an enzymatic method and carried out invitro.
• Polymerase chain reaction (PCR) is method for amplifying particular segments of DNA.
• Polymerase chain reaction (PCR) technique was developed by Kary mullis in the year of 1983.

PCR by OnlineBiotechNotes

PCR Steps

PCR consists of three basic steps.

1. Denaturation
2. Annealing
3. Extension

1. Denaturation:

• Two strands of DNA separate and form single stranded DNA.
• This step is carried out at 92C to 96C Temperature for 2 minutes.

2. Annealing:

• In Annealing, Primer to each strand is carried out at 45C to 55C Temperature.

3. Extension:

• DNA polymerase adds dNTPs (Deoxyribosenucleotide Triphosphate) which is complementary to templates strands at 3’end of primer.
• Extension is carried out at temperature of 72C.

Advantages of PCR

• PCR (polymerase chain reaction) is a simple and commonly used technique.
• It allows amplification of any specific sequence of DNA provided that short sequences either side of it are known.
• PCR Allow Rapid diagnosis and identification.

Applications of PCR

• PCR used in Forensic science: DNA finger printing, paternity testing and criminal identification
• PCR used in Diagnosis: Molecular identification of microorganisms
• PCR used in Evolution study: evolutionary biology
• PCR used in Fossil study: paleontology
• PCR used in Gene cloning and expression
• PCR used in Gene sequencing
• PCR used in Vaccine production by recombinant DNA technology
• PCR used in Drug discovery
• PCR used in Mutation study
• PCR used in Human genome project

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