1. Cutting of desired gene at specific location can be carried out by incubating purified DNA molecule with the restriction enzyme at some optimal condition. The progress of restriction enzyme digestion can be checked by Agarose gel electrophoresis.

2. Synthesis of multiple copies of the desired gene is also known as replication of DNA in vitro and can be achieved by PCR (Polymerase Chain Reaction). This involves three three main steps.

• Denaturation: The dsDNA is denatured by using high temperature of 95°C for 15 sec. Each separated ssDNA now acts as a template for the further synthesis of DNA.
• Annealing: This can be done by using two sets of oligonucleotide primers. This step is performed at somewhat lower temperature (40-60°C) using Mg²⁺ as a cofactor for Taq polymerase nd dNTPs depending upon the length and sequence of the primers.
• Extension: It is achieved by Thermostable enzyme, i.e. Taq polymerase in the reaction as it can tolerate the high amount of temperature and can extend the primers by adding nucleotides complimentary to the template.