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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.
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