List the three steps involved in Polymerase chain Reaction(PCR)

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Polymerase Chain Reaction (PCR)

  • Recombinant DNA can be amplified by PCR. Several identical copies of it can be synthesised in vitro.

  • Two sets of primers (chemically synthesised oligonucleotide stretches that are complementary to a region of DNA), enzyme DNA polymerase,and deoxynucleotides are added.

  • PCR consists of 3 steps:

    • Denaturation − Double helical DNA is denatured by providing high temperature. DNA polymerase does not get degraded in such high temperatures since the DNA polymerase used in this reaction is thermostable as it is isolated from thermophilic bacteria, Thermus aquaticus (Taq).

    • Annealing- It is the step in which primers are annealed to single stranded DNA templates.Two sets of primers (small chemically synthesised oligonucleotides that are
      complementary to the regions of DNA) are used. The temperature of reaction mixture is lowered to 50- 65°C for some seconds to allow annealing of primers. DNA polymerase extends the primer in 5' to 3' direction.
    • Extension − Replication of DNA occurs in vitro.

    • This cycle is repeated several times to generate up to 1 billion identical copies of the DNA.

       
          



Hope this information will clear your doubts about the polymerase chain reaction.
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