RECOMBINANT DNA TECHNOLOGY

Recombinant DNA is the general name for a piece of DNA that has been created by combining two or more fragments from different sources.

Process of Recombinant DNA Technology

The complete process of recombinant DNA technology includes multiple steps, maintained in a specific sequence to generate the desired product.

Step-1. Isolation of Genetic Material.

The first and the initial step in Recombinant DNA technology is to isolate the desired DNA in its pure form i.e. free from other macromolecules.

Step-2.Cutting the gene at the recognition sites.

The restriction enzymes play a major role in determining the location at which the desired gene is inserted into the vector genome. These reactions are called ‘restriction enzyme digestions’.

Step-3. Amplifying the gene copies through Polymerase chain reaction (PCR).

It is a process to amplify a single copy of DNA into thousands to millions of copies once the proper gene of interest has been cut using restriction enzymes.

Step-4. Ligation of DNA Molecules.

In this step of Ligation, the joining of the two pieces – a cut fragment of DNA and the vector together with the help of the enzyme DNA ligase.

Step-5. Insertion of Recombinant DNA Into Host.

In this step, the recombinant DNA is introduced into a recipient host cell. This process is termed as Transformation. Once the recombinant DNA is inserted into the host cell, it gets multiplied and is expressed in the form of the manufactured protein under optimal conditions.