Procedures of gene cloning

A clone refers to a group of organisms, cells or molecules arising from a single individual.The general procedures in gene cloning are:

1. Generation of desired DNA fragments:

via the methods of

• Restriction endonuclease digestion
• cDNA synthesis
• PCR 
• chemical synthesis

insertion of  fragments into a cloning vector

via :

• Ligation of blunt or cohesive ends 
• Homopolymer tailing 
• linker molecules 

Introduction into host cells

via:

• Transformation 
• transduction 
• transfection 

Selection or screening

via:

• Hybridisation 
• PCR 
• immunochemical methods 
• protein-protein interactions 
• functionl complementation 

Flowchart of gene cloning procedure