Question

Mention the different steps of the process of recombinant DNA technology?

Answer 1

Recombinant DNA technology (rDNA) means the association of DNA molecules from two different species that are embedded into a host to create a new genetic combination that is of worth to science, medicine, horticulture, and industry. There are six steps that are involved in recombinant DNA technology.

Complete answer:
-Isolation of genetic material: The initial step in this technology is to isolate the desired DNA in its pure form means free from other macromolecules like RNA, polysaccharides, proteins, and lipids, with the help of enzymes such as lysozymes, ribonuclease, etc.
-Restriction enzyme digestion: Restriction enzyme such as endonuclease is known as molecular scissors because they cut the DNA at the desired place. They include the incubation of the DNA with the selected restriction enzyme at conditions ideal for that enzyme.
-Amplification of DNA: Polymerase chain reaction is a technique used for the amplification of desired DNA. It helps to amplify single DNA into thousands of copies of DNA.
Ligation of DNA molecule: The fragments of desired DNA and cloning vector are cut with the same restriction enzyme and the process of joining both pieces together by using enzyme DNA ligase are known as DNA ligation.
-Insertion of Recombinant DNA: Recombinant DNA is inserted into the host cell and the process is known as transformation.
-Isolation of recombinant cells: A process of transformation generates both transformed and non-transformed host cells. Isolation of recombinant cells from non-recombinant cells is occurred by using selectable marker genes of the plasmid vector.

Note: A DNA that undergoes amplification is known as Template.PBR322 plasmid vector contains a distinct selectable marker gene i.e. Ampicillin and Tetracycline gene. A small part of tissue contains many kilometres of DNA.