The various steps involved in gene cloning technique are :
• Identification and isolation of desired DNA. .
• Amplification of gene of interest using PCR.
• Fragmentation/cutting of desired DNA and vector DNA by restriction enzymes.
• Ligation/joining of desired DNA fragment into vector using ligase enzyme.
• Transferring the recombinant DNA into host cell/organism by transformation, transfection, electroporation, microinjection (vectorless transfer), particle bombardment gun (Biolistics) (Vectorless transfer) or by Agrobacterium/retrovirus mediated gene transfer..
• Culturing the host cell for obtaining the foreign gene product/recombinant protein.
• Extraction of desired products or downstream processing.
