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Processes of Recombinant DNA Technology. ( e - Ratpl.o A, Recombinant DŇA technology involves the following steps :, (i) Isolation of DNA., (ii) Fragmentation of DNA by restriction endonucleaseş.., Li Isolation of the desired-DNA fragment( separation and isolation oF DNA fragmes, (iv) Amplification of the gene of interest. using PCR, (v) Ligation of the DNA fragment into a vector using DNA ligase., (vi) Transfer of recombinant DNA into the host., (vii) Culturing the host cells on a suitable medium on a large scale., (viii) Extraction of the desired product., (ix) Downstream processing õf the product as a finished product ready for marketing., (), Isolation of DNA C ISolation R genetc material (DNA), 70ct16, DNA has to be isolated in pure form for the action of restriction enzymes., DNA can be released from the cells by digesting the cell envelope by the use of enzymes, like lysozyme for bacterial cells, chitinase for fungal cells and cellulase, for plant cellsf alssras., Since DNA is intertwined with histone proteins and RNA proteins are removed by treatment, with proteases and RNAS by ribonucleases., Other impurities are removed by employing suitable treatments., The purified DNA is precipitated by the addition of chilled ethanol; it is seen as fine threads, in suspension., wined, स्वासंनई, -, लपेघना, (ii) Fragmentation DNA., गुती,, Fragmentation of DNA is carried out by incubating the purified. DNA molecules with suitáble, restriction enzymes at optimal conditions of temperature and pH., ANA