Protocols.io also provides an interactive version of this protocol where you can discover and share optimizations with the research community. It is available for both the Non-Radioactive Phosphorylation with T4 PNK and the Non-Radioactive Phosphorylation with T4 PNK 3' phosphatase minus.
Set-up the following reaction:
||up to 300pmol of 5' termini
|10X T4PNK Reaction Buffer
||1 µl (10 units)
||up to 50 µl
Incubate at 37°C for 30 minutes.
Heat inactivate by incubating at 65°C for 20 minutes.
- ATP is not supplied
- 1X T4 DNA Ligase Buffer contains 1 mM ATP and can be substituted in non-radioactive phosphorylations (T4 Polynucleotide Kinase exhibits 100% activity in this buffer).