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Star Republic: Guide for Biologists

Terminal deoxynucleotidyl transferase

1. Set up the transferase reaction:

DNA (0.1-1 mg, or ~1 pmol) 4 ml
100 mM dNTP 2 ml
5X Reaction Buffer
(1 M potassium or sodium cacodylate, 125 mM Tris-HCl pH 7.2,
0.05% Triton X-100 and 5 mM CoCl2)
4 ml
Terminal Deoxynucleotidyl Transferase (20 units/ml) 2 ml
ddH2O 8 ml
Total volume 20 ml

2. Incubate at 37 C for 10-30 minutes.

3. Heat inactivate at 65 C for 10 minutes or add 2 ml 0.5 M EDTA.

See also vector for TA cloning.