Process of Recombinant DNA Technology : Isolation of DNA =>Cutting of DNA using restriction endonuclease => Amplification of Gene using PCR => Making rDNA and insertion of it into host cell/organism => obtaining the foreign gene product => Downstream processing.
(i) Isolation of Genetic Material (DNA) :
DNA can be obtained from the cell by treating with enzymes like, Lysozyme for bacteria, Cellulase for plant cell, Chitinase for fungus.
- Histone protein and RNA can be removed by treating with proteases and ribonuclease respectively.
- Purified DNA precipitated by the addition of chilled ethanol, fine threads of DNA are obtained in the suspension.
