Molecular cloning is the first laboratory step in producing a recombinant protein. The aim is to obtain a plasmid that carries the gene of interest (GOI) in an expression vector.

A general methodology is described briefly below:

  1. Obtain/generate a DNA template of the GOI
  2. Ligate the GOI into an appropriate expression vector (ligation dependent cloning)
  3. Transform the expression vector into a bacterial strain
  4. Analyse bacterial clones to confirm integration of the GOI
  5. Sequence verification