Mediating the recombination reaction

As explained, the core of Invitrogen™ Gateway™ cloning is the Entry vector. Once the Entry clone is ready, the gene of interest is easily shuttled to a secondary plasmid, the Destination vector. This reaction is mediated by a robust enzyme mixture called LR Clonase™, which contains the necessary protein activity to excise the gene of interest from the Entry clone and integrate it into the Destination vector, which then becomes your expression clone (Figure 1). Reversing this reaction simply entails performing a BP reaction with BP Clonase enzyme mix. Both LR and BP Clonase enzyme mixtures are easy-to-use master mix formats ensuring consistency and reliability from reaction to reaction.

Ordering information

Types of Clonase enzymes

Gateway BP Clonase Enzyme Mixes

Gateway BP Clonase enzyme contains both Int (Integrase) and IHF (Integration Host Factor) proteins that catalyze the in vitro recombination of PCR products or DNA segments from clones (containing attB sites) and a Donor vector (containing attP sites) to generate Entry clones. View the selection guide below to find the best BP Clonase™ enzyme for your research goals.

Gateway LR Clonase Enzyme Mixes

Gateway LR Clonase II enzyme mix contains a proprietary blend of Int (Integrase),  IHF (Integration Host Factor) and Xis (Excisionase) enzymes that catalyze the in vitro recombination between an Entry clone (containing a gene of interest flanked by attL sites) and a Destination vector (containing attR sites) to generate your expression clone. We offer different formats of LR Clonase enzyme mixtures, depending on your application and desired format. View the selection guide below to find the best LR Clonase enzyme for your research goals.

Figure 1. The Gateway reactions.
 

Clonase Enzyme Selection Guide

Rating Enzyme Format Advantages
LR Clonase Enzyme Mixes
Best LR Clonase II Plus Enzyme Mix Single mix of enzyme and reaction buffer
  • Highest cloning efficiency for single and multiple fragment cloning
  • Optimized for difficult cloning reactions
  • Works with MultiSite Gateway Pro Technology
  • Easy to use single mix format ensures enzyme stability
  • Convenient 10 µl reaction set up
Very good LR Clonase II Enzyme Mix Single mix of enzyme and reaction buffer
  • High efficiency for single fragment cloning
  • Convenient master single mix format eliminates pipetting steps and error
  • Easy to use single mix format ensures enzyme stability
  • Convenient 10 µl reaction set up
Good LR Clonase Enzyme Enzyme
5X reaction buffer
  • Basic cloning of only single fragments
BP Clonase Enzyme Mixes
Best BP Clonase II Enzyme Mix Single mix of enzyme and reaction buffer
  • High efficiency cloning for Entry Clone construction
  • Single mix format eliminates pipetting steps and error
  • Easy to use single mix format ensures enzyme stability
  • Convenient 10µl reaction set up
Good BP Clonase Enzyme Enzyme
5X reaction buffer
  • Basic cloning of only single fragments
Ordering information