Question

Explain the various steps in recombinant technology.

Answer 1

The technology used for cutting and joining of DNA molecules belonging to two different species is called biotechnology. It takes place in order to obtain the gene of interest. The main steps involved in recombinant technology are isolation of the genetic material, restriction enzyme digestion, amplification, ligation, and Inserting the recombinant DNA into the host.

Complete answer: The basic steps involved in recombinant DNA technology are:
1. Identification of DNA with desirable genes.
2. Introduction of identified DNA into a suitable host to form recombinant DNA.
3. Maintenance of the introduced DNA in the host and transfer the DNA to its progeny.
a) Isolation of genetic material is the initial step involved in the process. The DNA is separated and purified with the help of enzymes such as lysozymes, cellulase, chitinase, protease, ribonuclease, etc
b) The DNA is cut at specific locations using molecular scissors called restriction enzymes. This process is called restriction enzyme digestion. The purified DNA is incubated with selected restriction enzymes. For revealing the progress of restriction enzyme digestion a process called agarose gel electrophoresis is carried out. In agarose gel electrophoresis, it involves running the DNA fragments on an agarose gel with the application of electricity. The negatively charged DNA moves towards the positive electrode and is separated out on the basis of size.
c) Then the polymerase chain reaction is carried out for amplifying a single copy or few numbers of copies into thousand to millions of copies.
d) Then the ligation of the DNA is carried out using the enzyme DNA ligase. The processes in which two DNA fragments are joined together are known as ligation.
e) Then the process of transformation is carried out. The process of introducing the recombinant DNA into a host organism is called transformation. Before transformation DNA must be made competent. Thermal shocking, Calcium ion treatment, electroporation, etc. is carried out to make DNA competent.
f) As a result of the transformation, a mixed population of transformed and non transformed host cells are formed. Later transformed cells are filtered out.

Note: Recombinant DNA technology is most commonly used in biotechnology, medicine, and research. This technology was developed by Stanley Cohen. Biotechnology mainly uses plasmids as vectors.