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进口试剂
进口试剂
T4 Polynucleotide Kinase
T4 Polynucleotide Kinase
- Recombinant T4 Polynucleotide Kinase (T4 PNK).
- Applications include end-labeling and insert preparation for cloning.
T4 Polynucleotide Kinase catalyzes the transfer and exchange of gamma-phosphate of ATP to the 5´-hydroxyl terminus of polynucleotides (double-and single-stranded DNA and RNA) and nucleoside 3´-monophosphates.
Source:
Purified from a strain of E. coli that carries a T4 Polynucleotide Kinase expression plasmid.
Applications:
- End-labeling DNA for probes and DNA sequencing (1)
- Addition of 5´-phosphates to oligonucleotides to allow subsequent ligation (2)
Heat inactivation: 70°C for 15 minutes.
Storage:
Enzyme and buffer should be stored at -20°C. Enzyme is stable for 12 months if handled properly.
Additional Information:
- Ammonium ions are potent inhibitors of T4 Polynucleotide Kinase. DNA should not be precipitated from or dissolved in buffers containing ammonium salts.
- Protruding 5’-overhangs are more efficiently phosphorylated than blunt- or recessed 5’ overhangs.
References:
- Sambrook, J. and Russell, W. (2001) Molecular Cloning: A Laboratory Manual, (3rd ed.), 9.55-9.56, 9.68-9.69.
- Sambrook, J. and Russell, W. (2001) Molecular Cloning: A Laboratory Manual, (3rd ed.), 13.16-13.17.
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