T4 Polynucleotide Kinase

Downloads

Certificate of Analysis PDF -Current Lot
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Reagents Supplied

10X T4 Polynucleotide Kinase Reaction Buffer

Applications

• Catalyzes the transfer of the gamma-phosphate of ATP to the 5'-OH terminus in DNA or RNA
• Contains 3'-phosphatase activity (1)
• Used for 5'-end labeling of nucleic acids prior to DNA or RNA sequencing (2, 3)
• Phosphorylates synthetic linkers and fragments of DNA or RNA prior to ligation
• Labels 5'-termini prior to partial restriction enzyme digestion

Unit Definition

One unit is the amount of enzyme required to transfer 1 nmol of γ-phosphate from ATP to the 5' OH terminus of salmon sperm DNA at 37°C in 30 minutes

Heat Inactivation

15 minutes at 65ºC

Concentration

20 - 30 Units/µl

Storage Buffer

50 mM Tris HCl (pH 7.6)
0.1 mM EDTA
25 mM KCl
1 mM dithiothreitol
0.1 µM ATP
50% glycerol

Assay Conditions

70 mM Tris-HCl (pH 7.6) 
10 mM MgCl₂
5 mM dithiothreitol
17.4 µg salmon sperm DNA
70 mM [γ-³²P] ATP 
Reaction volume of 100 µl

Storage Conditions

Store at -20°C
Product shipped on dry ice

References

(1) Richardson, C.C. (1971) Progress in Nucleic Acids Res. and Mol. Biol. 2, 815-828
(2) Maxam, A. and Gilbert, W. (1977) Proc. Natl. Acad. Sci. USA 74, 560-56
(3) Donis-Keller, H. (1980) Nucleic Acids Res. 8, 3133-3142