Modification of the termini of double-stranded DNA is often necessary to prepare the molecule for cloning. DNA ligases require a 5' monophosphate for adenylation of the donor end, while the acceptor end requires a 3' hydroxyl group. Additionally, the sequences to be joined need to be compatible: either a blunt end being joined to another blunt end, or a cohesive end with a complementary overhang to another cohesive end. End modifications are performed to improve the efficiency of the cloning process, ensure the ends to be joined are compatible, and to optimize the positioning of regulatory and translated sequences.
Molecular Cloning Technical Guide
Download the latest Molecular Cloning Technical Guide for help with product selection, protocols, tips for optimization and trouble-shooting.
Reagents & Tools for Molecular Cloning brochure
Learn about recommended products for cloning in our Reagents and Tools for Molecular Cloning Brochure.
- A-tailing Selection Chart
- Blunting Selection Chart
- DNA Ligase Selection Chart
- Phosphatase Selection Chart
- Troubleshooting Guide for Cloning
- Troubleshooting Guide for End Modification
- Troubleshooting Tips for Ligation Reactions
- Tips for Maximizing Ligation Efficiencies
- Traditional Cloning Quick Guide