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. T4Polynucleotide 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. To learn more about how to identify what type of overhang you have, visit this video tutorial. 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
This product is related to the following categories:
DNA Manipulation Products
This product can be used in the following applications:
Blunting,
Phosphorylation (Kinase),
PCR,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB