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
Applications:
-
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 DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
Supplied with:
10x T4 Polynucleotide Kinase Buffer
10xT4 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 Condition: -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.
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