At 52 C primer hybridization will take place at which step
a) Denaturation
c) Amplification
b) Annealing
d) None of the abo
Answers
Answer:
At the start of PCR, the DNA from which a segment is to be amplified, an excess of the two primer molecules, the four deoxynucleoside triphosphates and the DNA polymerase are mixed in the reaction mixture that has appropriate quantities of Mg2+.
The reaction mixture is first heated to a temperature between 90−98oC (commonly 94oC) that ensures DNA denaturation. Every single strand of the target DNA then acts as a template for DNA synthesis. This is the denaturation step. The duration of this step in the first cycle of PCR is usually 2 min at 94oC.
The mixture is now cooled to a temperature (generally 40−60oC) that permits annealing of the primer to the complementary sequences in the DNA. This step is called annealing. The duration of the annealing step is usually 1 min.
The primers are extended towards each other so that the DNA segment lying between the two primers is copied, this is ensured by employing primers complementary to the 3': ends of the segment to be amplified. The duration of primer extension is usually 2 min at 72oC.