The T4 Polynucleotide Kinase (NEB #M0201) can be used to 5' end label DNA and RNA because it catalyzes the transfer and exchange of phosphate from ATP to the 5´ -hydroxyl terminus of polynucleotides (double-and single-stranded DNA and RNA) and nucleoside 3´-monophosphates. Polynucleotide Kinase also catalyzes the removal of 3´-phosphoryl groups from 3´-phosphoryl polynucleotides, deoxynucleoside 3´-monophosphates and deoxynucleoside 3´-diphosphates (1).
The T4 Polynucleotide Kinase (3' phosphatase minus) (NEB #M0236) is a modified version that exhibits full kinase activity with no 3´ phosphatase activity (2,3).
NEBCloner is a guide for selecting appropriate products and viewing protocols for steps in the cloning workflow. To help select the right kinase, choose "End Modification" on the tool to start.
- Richardson, C.C. (1981) P.D. Boyer (Eds.), The Enzymes, 14, pp. 229-314. San Diego: Academic press.
- Wang, L.K. and Shuman, S. (2001) J. Biol. Chem., 276, 26868-26874. PMID: 11335730
- Wang, L.K. and Shuman, S. (2002) Nucl. Acids Res., 30, 1073-1080. PMID: 11842120