Description
T4 Polynucleotide Kinase (PNK) catalyzes the transfer and exchange of the terminal gamma position phosphate of ATP to the 5′-hydroxyl terminus of double-and single-stranded DNA, RNA and nucleoside 3′-monophosphate molecules (1).
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Description: T4 Polynucleotide Kinase (PNK) catalyzes the transfer and exchange of the terminal gamma position phosphate of ATP to the 5'-hydroxyl terminus of double- and single-stranded DNA, RNA and nucleoside 3'-monophosphate molecules(1). T4 PNK also exhibits 3'-phosphatase and 2',3'cyclic phosphodiesterase activities.(2-6) Specific Activity: 133,333 U/mg Application: - End-labeling DNA or RNA for probes and DNA sequencing - Addition of 5'-phosphates to oligonucleotides to allow subsequent ligation - Removal of 3'-phosphoryl groups Source: Purified from a strain of E. coli that expresses the recombinant T4 Polynucleotide Kinase gene. Supplied in: 10 mM Tris-HCL 50 mM KCl 0.1 µM ATP 1.0 mM Dithiothreitol 0.1 mM EDTA 50% Glycerol pH 7.4 @ 25°C Supplied with: 10x T4 Polynucleotide Kinase Buffer 10x Polynucleotide Kinase Buffer: 700 mM Tris-HCL 100 mM MgCl2 50 mM DTT pH 7.6 @ 25°C Unit Definition: One unit is defined as the amount of enzyme catalyzing the incorporation of 1 nmol of [32P] (ATP donor) in 30 minutes at 37°C in 1x T4 Polynucleotide Kinase Reaction Buffer. Recommended Storage Conditions: -20°C Reference: 1. Richardson, C.C. (1981) P.D. Boyer (Eds.), The Enzymes, 14, pp. 229-314. San Diego: Academic press. 2. Morse, D. P. et al. (1997) Biochemistry 36, 8429-8434. 3. Cameron, V. et al. (1977) Biochemistry 16, 5120-5126. 4. Wand, L. K. et al. (2002) Nucl. Acids Res. 30, 1073-1080. 5. Galburt, E., et al. (2002) Structure 10, 1249-1260. 6. Wang, L. K., et al. (2002) EMBO J. 21, 3873-3880. |
