Isolated from E.coli strain that carries the cloned Polynucleotide kinase gene from bacteriophage T4
Description
T4 polynucleotide Kinase catalyzes the transfer and exchange of Pi from the γ position of ATP to the 5'hydroxyl terminus of polynucleotides (double- and single-stranded DNA and RNA) and nucleoside 3'monophosphates. The enzyme also catalyzes the removal of 3'- phosphoryl groups from 3'- phosphoril polynucleotides, deoxynucleoside 3'- monophosphates and deoxynucleoside 3'- diphosphates.
Applications:
- end-labeling DNA or RNA for probes and DNA sequencing;
- addition of 5'- phosphates to oligonucleotides to allow subsequent ligation;
- removal of 3'- phosphoril groups.
Unit
One unit is the amount of enzyme catalyzing the incorporation of 1 nmol of acid-insoluble [32P] in 30 minutes at 37oC.
Unit Assay Conditions: 1 x T4 polynucleotide kinase Buffer, 66 uM [γ- 32P] ATP (5 x 106 cpm/umol) and 0.26 mM 5'- hydroxyl - terminated salmon sperm DNA.
Reaction buffer
SE-buffer polynucleotide kinase T4, (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 5 mM DTT.)
Optimal temperature
37oC
Storage conditions
10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 0.1 mM EDTA; 1 mM DTT; 50% glycerol; Store at -20oC.
Quality control
Free of exonuclease, phosphatase, endonuclease and RNase activities
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