Polymerase chain reaction or PCR uses repeated cycles of heating and cooling to make the copies of specific DNA. High temperature is necessary to break weak hydrogen bond that binds the two stands of DNA together and as a result multiple copies of a specific DNA sequence can be obtained.
In order to carry out PCR successful, following components are required.
Target DNA molecule,
A pair of DNA primers,
Heat resistant DNA polymerase,
All four types of deoxynucleotide triphosphate (dNTPs)
Target DNA molecule is a segment of double stranded DNA that contains the …show more content…
Denaturation ( DNA strand separation)
Annealing ( Hybridization of primers)
Extension ( DNA synthesis, elongation,replication)
Step One: This is the first step of denaturation in which the double stranded DNA is heated to about 95 degree Celsius for about 15 seconds. Temperature is increased to break down the hydrogen bond that binds the double strands of DNA together to split into two single stands of DNA.
Step two: Once the two strands of DNA are separated , Temperature is reduced to 54 degree Celsius to cool down the mixture solution. DNA primers will begin to bind the flanking sequence. DNA primers binds to the 3 end of one strand and another primer binds to the 3 end of the complementary strand.
Step three: Deoxynucleotide triphosphate (dNTPs) and temperature resistant polymers are added in this elongation step. The temperature is slightly increased to 72 degree Celsius as it is the optimal temperature of heat resistant DNA polymerase