Radioactive Labeling with T4 PNK 3' phosphatase minus (M0236)
New England Biolabs
Published: 2022-02-20 DOI: 10.17504/protocols.io.bd23i8gn
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Abstract
Radioactive Labeling with T4 Polynucleotide Kinase 3' phosphatase minus.
Before start
Steps
1.
Note: [33P] ATP may be substituted for [32P] ATP.
Set-up the following reaction:
| A | B |
|---|---|
| Component | Amount |
| DNA | Up to 50 pmol of 5´ termini |
| T4 PNK Reaction Buffer (10X) | 5 µl |
| ATP (10 mM) | 50 pmol of [γ-32P] ATP |
| T4 PNK | 2 µl (20 units) |
| Nuclease-free Water | up to 50 µl |
Note
2.
Incubate at 37°C for 0h 30m 0s.
3.
Heat inactivate by incubating at 65°C for 0h 20m 0s.
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