Describe the process of gene amplification for rDNA technology experiments.

Amplification of a gene sample of interest is carried out using PCR in the following three steps:

1. Denaturation-

•The double-stranded DNA is denatured by applying a high temperature of about 95 degree Celsius for 15 seconds.

•Each separated single-stranded strand now acts as a template for the synthesis of a new DNA strand.

2. Annealing-

•Two sets of primers are added which anneal at the 3’ end of each separated strand. They mark the beginning of replication.

•It occurs at about 40 to 60 degree Celsius depending on the length and sequence of primers.

3. Extension

•A thermostable DNA polymerase, Taq polymerase extends the primers by adding nucleotides complementary to the template provided in the reaction.

•Polymerisation is usually carried out at 72 degree Celsius.