E1201S, Quick Blunting Kit - 20 reactions

The Quick Blunting Kit is used to convert DNA with incompatible 5´ or 3´ overhangs to 5´ phosphorylated, blunt-ended DNA for efficient blunt-end ligation into DNA cloning vectors. DNA is blunted using T4 DNA Polymerase (NEB #M0203) which has both 3´ → 5´ exonuclease activity and 5´ → 3´ polymerase activity. T4 Polynucleotide Kinase (NEB #M0201) is included in the enzyme mix for phosphorylation of the 5´ ends of blunt-ended DNA for subsequent ligation into a cloning vector. This kit is optimized for blunting up to 5 µg of DNA in a single reaction.

Blunt Enzyme Mix supplied in: 100 mM KCl, 10 mM Tris-HCl(pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 0.1% Triton X-100 and 50% Glycerol.

1X Blunting Buffer:
100 mM Tris-HCl
50 mM NaCl
10 mM MgCl₂
0.025% Triton X-100
5 mM dithiothreitol
pH 7.5 at 25°C