The Ligase Experts
Overview T4 DNA Ligase Comparison Troubleshooting Tips for
Ligation Reactions


  • Add controls, including vector alone, insert alone, and uncut vector
     
  • Vary the molar ratio from 1:1 to 1:5 vector:insert
     
  • I nsert or plasmid should have a 5´ phosphate
     
  • Use fresh buffer as the ATP or DTT may degrade over time
     
  • Total DNA concentration should be less than 10 ng/μl
     
  • When excising a fragment from a gel, minimize UV exposure and use longwave UV (360 nm) to reduce damage to the ends of the DNA