Polynucleotide Kinase T4
Catalog Number:
USB-P4410
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| Article Name: | Polynucleotide Kinase T4 |
| Biozol Catalog Number: | USB-P4410 |
| Supplier Catalog Number: | P4410 |
| Alternative Catalog Number: | USB-P4410-500,USB-P4410-2500 |
| Manufacturer: | US Biological |
| Category: | Molekularbiologie |
| T4 polynucleotide kinase catalyzes the transfer of the terminal phosphate of ATP (gamma-orthophosphate) to the 5-hydroxyl termini of ribo- and deoxyribonucleotides. In the presence of ADP the enzyme also catalyzes an exchange reaction. In the exchange reaction, an excess ADP causes the enzyme to transfer the 5 terminal phosphate from phosphorylated DNA to ADP. The DNA thus dephosphorylated is then rephosphorylated by transfer of radiolabeled gamma-phosphate from gamma 32P ATP. Applications: 5 Labeling of nucleic acids - probes for hybridization - probes for transcript mapping - markers for gel electrophoresis - primers for DNA sequencing - primers for PCR 5phosphorylation of oligonucleotides, PCR products, other DNA or RNA prior to ligation Phosphorylation of PCR primers Detection of DNA modification by the [32P]-postlabeling assay Removal of 3-phosphate groups Source: E. coli infected with T4 phage Form: Supplied as a liquid in 20mM Tris-HCl, pH 7.5, 25mM KCl, 0.1mM EDTA, 2mM DTT, 50% glycerol,. Concentration: 10u/ul Unit Definition: One unit will incorporate 1 nanomole of gamma phosphate from ATP to 5-OH DNA in 30 minutes at 37C. Activity Assay Buffer: 100mM Tris-HCl, pH 8.0, 10mM MgCl2, 5mM DTT, 0.5mM 5-OH DNA, 0.05mM ATP, 0.1MBq/ml [g33p]-ATP. Inhibitors: Metal chelators, phosphate and ammonium ions, KCl and sodium chloride at a concentration higher than 50mM. Inactivation: Heat at 75C for 10 min or by the addition of EDTA. Supplied with: P4410A: Forward Reaction Buffer (10X): Supplied as a liquid in 500mM Tris-HCl, pH 7.6,n100mM MgCl2, 50mM DTT, 1mM Spermidine. P4410B,, Exchange Reaction Buffer (10X): Supplied as a liquid in 500mM Imidazole-HCl, pH 6.4, 180mM MgCl2, 50mM DTT, 1mM Spermidine, 1mM ADP. P4410C: PEG Solution: Supplied as a liquid, 24% w/v PEG 6000. Quality Control: 1. Endodeoxyribonuclease Assay: No detectable degradation was observed after incubation of supercoiled plasmid DNA with T4 Polynucleotide Kinase. 2. Ribonuclease Assay: No detectable degradation was observed after incubation of [3H] with T4 Polynucleotide Kinase. Storage and Stability: May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
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