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Explain the process of amplification of genes of interest using PCR technique.

 

 

 

 
 
 
 
 

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PCR ( Polymerase chain reaction) involves three steps

  1.  Denaturation- The double stranded DNA is heated so that the two strands separate from each other (Temp- 94^{\circ}C)
  2.  Annealing- The primers which are small chemically synthesized oligonucleotides that are complementary to the region of DNA ( our gene of interest) attach to the template DNA. ( Temp- 50-55^{\circ}C)
  3.  Extension- The enzyme DNA Polymerase extends the primers using the nucleotides provided in the reaction and the genomic DNA as template. ( Temp- 74^{\circ}C) The process of replication of DNA is repeated many times, the segment of DNA can be amplified to approximately billion times, i.e., 1 billion copies are made. Such repeated amplification is achieved by the use of a thermostable DNA polymerase (isolated from a bacterium, Thermus aquaticus), which remain active during the high temperature.

 

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Priyanka Kumari

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