DNA End Repair Mix offers an easy, one step process for creating blunt ends in sheared, sonicated, or restriction-cut DNA and PCR fragments for efficient ligation into blunt end cloning vectors.DNA End Repair Mix allows you to:
A Simple Way to Make Blunt Ends
- Generate blunt ends in DNA and PCR fragments (A-tail)
- Repair DNA with 5′ or 3′ overhangs in 30 minutes at 37 °C
- Heat inactivate DNA End Repair Mix in 10 minutes at 75 °C
With DNA End Repair Mix, DNA with 5′ or 3′ overhangs is converted to 5′ phosphorylated blunt end DNA in 30 minutes at 37 °C. Once DNA has been repaired, the enzymes in DNA End Repair Mix can be completely inactivated in 10 minutes at 75 °C thus preventing similar fill-in or repair of DNA subsequently added to the mix.
DNA End Repair Mix enzymes are premixed to minimize pipetting. T4 DNA polymerase, with both 3′ -> 5′ exonuclease activity and 5′ -> 3′ polymerase activity is optimized for filling or cutting 5′ or 3′ overhangs. T4 polynucleotide kinase is included for phosphorylation of 5′ ends. DNA End Repair Mix is optimized for blunting up to 5 μg of DNA at a time.DNA End Repair Mix Buffers:
- 10X DNA End Repair Mix Buffer: 500 mM Tris-HCl (pH 7.5); 100 mM MgCl2, 100 mM DTT, 10 mM ATP, 4 mM dATP, 4 mM dCTP, 4 mM dGTP, 4 mM dTTP.
- DNA End Repair Enzyme Mix Storage Buffer: 10 mM Tris-HCl (pH 7.5); 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol, 0.1% Triton® X-100.
- 10X DNA End Repair Enzyme Mix: 20 reactions
- 10X DNA End Repair Reaction Buffer: 200 μl