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1. T4 polynucleotide kinase can be used to label double-stranded
and single-stranded DNA or RNA at the 5'-ends.
2. g-32P-dATP is highly
radioactive. Proper protection and waste disposal are required.
3. Prepare the following mixture:
| ddH2O |
4 ml |
| 10X T4 polynucleotide kinase buffer |
1 ml |
| DNA (0.1-1mg, 50-200 ng oligos) |
2 ml |
| g-32P-dATP (5-10 gCi) |
2 ml |
| T4 polynucleotide kinase |
1 ml |
| Total |
10 ml |
4. Incubate at 37 °C in a heat block reserved for radioactive reactions for 30-60 minutes.
5. Heat inactivate the T4 polynucleotide kinase at 65 °C for 10 minutes.
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