Directional cloning method requires so many steps with less efficiency. It is also time-consuming. Before cloning a gene of interest it is mandatory to have a common restriction site between the vector backbone and on the flanks of the gene of interest. Otherwise, it is quite impossible to clone. However, this constraint can be overcome with the help of adapter or linker molecules.

Whereas the efficacy of gateway cloning is over 90% and once the Entry vector is prepared the gene of interest could be cloned into Destination vector for developing Expression Vector in just one step within an hour only. Moreover, this doesn’t require having common restriction sites between the plasmid vector and the gene of interest as the earlier mentioned method.

Pros of Gateway cloning:

  • Very precise method of cloning
  • Time-saving protocol
  • No restriction digestion is required

Cons of Gateway Cloning:

  • Requires costly reagents, e.g. BP Clonase, LR Clonase
  • Specific vectors and strains are required
  • Initial set up is time-consuming

Prepared by
Aqief Afzal
Research Assistant