Polynucleotide Kinase from T4-infected Escherichia coli
SIGMA/P4390 - 10 units/μL, buffered aqueous glycerol solution
CAS Number: 37211-65-7
MDL Number: MFCD00166009
Product Type: Chemical
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| concentration | 10 units/μL |
| foreign activity | Endonuclease and exonuclease, none detected |
| form | buffered aqueous glycerol solution |
| grade | Molecular Biology |
| mol wt | 33 kDa |
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | −20°C |
| Analysis Note: | Activity is determined in a reaction mixture containing 40 mM Tris-HCl (pH 7.5), with 10 mM MgCl2, 5 mM dithiothreitol, 0.5 mM 5’-OH polynucleotide ends, and mM [γ-32P]-ATP. |
| Application: | Suitable for: • Sequencing or nucleic acid tagging (DNA and RNA) by 5′-end labeling • 5′ phosphorylation of oligonucleotides • Removal of 3′-phosphate groups from phosphorylpolynucleotides |
| Biochem/physiol Actions: | Polynucleotide kinase catalyses a "forward reaction" transfer of the γ-phosphate of ATP to the 5′ hydroxyl terminus of single- and double-stranded nucleic acids (DNA and RNA) and 3′-nucleoside monophosphates. In exchange reactions containing ADP, the enzyme will catalyze the exchange of 5′-terminal phosphate groups and ATP. The 3′-phosphatase activity enables the enzyme to remove 3′-phosphoryl groups from phosphorylpolynucleotides 1. Forward reaction: Transfer of the labeled γ-phosphate from [γ-32P]-ATP to the free 5′-hydroxyl group of the substrate. 5′-HO-DNA + [γ-32P]-ATP → 5′-32PO-DNA + ADP. Substrates that do not have a free 5′-hydroxyl require prior dephosphorylation by alkaline phosphatase. 2. Exchange reaction: First, the terminal 5′-phosphate is transferred from the substrate to ADP present in the reaction mixture. Then, the labeled γ-phosphate from [γ-32P]-ATP is transferred to the free hydroxyl group of the substrate. 5′-PO-DNA + ADP → 5′-HO-DNA + ATP 5′-HO-DNA + [γ-32P]-ATP → 5′-32PO-DNA + ADP |
| Other Notes: | One unit catalyzes the transfer of one nanomole of 32P to the 5′-end of micrococcal nuclease-treated DNA in 30 min. at 37 °C. Transfer is detected as incorporation into acid-insoluble material. |
| Other Notes: | T4 Polynucleotide Kinase is supplied in a solution of 50% glycerol (v/v), 20 mM Tris-HCl (pH 7.5), 25 mM KCl, 2mM DTT, 0.1 mM EDTA, and 0.1 μM ATP. |
| Symbol |  GHS08 |
| Signal word | Danger |
| Hazard statements | H334 |
| Precautionary statements | P261 - P284 - P501 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| Enzyme Commission (EC) Number | 2.7.1.78 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |