Describe the various steps involved in Recombinant DNA technology
The steps involved in the recombinant DNA are: i) Identification of DNA with Desirable Genes: The addition of chilled ethanol will result in obtaining the purified DNA while by using various other appropriate techniques the other molecules in the target cell can also be removed. ii) Cutting the DNA at a Specific Location: The source and vector DNAS after being cut will be introduced together having a gene of interest and specific restriction site and will be joined together with the help of an enzyme called ligase. iii) Insertion of Recombinant DNA into Host Cell: The bacterial cell is first treated with the specific concentration of divalent cation e.g., Ca2+ so that they can be competent enough to take up the DNA in the plasmid. The calcium ions increase the efficiency of DNA to enter into a bacterium through pores in its cell wall. It is a vector-less method. iv) Selection & Screening: The selectable marker for this recombinant plasmid will be the ampicillin resistance gene. They after placing them on an ampicillin-containing medium will undergo the process of plating that will help in the differentiation between the trAns.formants from non-trAns.formants. v) Obtaining the Foreign Gene product: The gene of interest after its cloning along with the presence of optimum growth conditions will result in the expression of target proteins which then needs to be produced on a large scale.
