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| Product Name | T4 Polynucleotide Kinase for 5 Prime Phosphorylation and End Labeling (10 U/uL) |
| Catalog No. | IVDRM-0460 |
| Description | T4 polynucleotide kinase transfers the gamma phosphate of ATP to the 5 prime hydroxyl end of oligonucleotides, whether single or double stranded DNA or RNA, and the reaction is reversible. The enzyme also carries a 3 prime phosphatase activity that removes 3 prime phosphate groups from oligonucleotides, deoxy 3 prime monophosphates and deoxy 3 prime diphosphates, and in the presence of ADP it exhibits 5 prime phosphatase activity that exchanges 5 prime phosphate groups with ATP. Typical workflows include phosphorylating DNA or RNA ends for ligation, end labeling for probes and DNA sequencing, preparing pNp substrates for 3 prime tagging, and labeling oligonucleotides whose 3 prime ends carry phosphates. Magnesium and sulfhydryl reagents such as DTT serve as cofactors, while phosphate, pyrophosphate and sodium phosphate inhibit activity. |
| Intended Use | 5 prime phosphorylation for ligation, end labeling for probes and sequencing, pNp substrate preparation and NGS adapter workflows |
| Principle / Technology | ATP gamma phosphate transfer to 5 prime hydroxyl ends with reversible kinetics and 3 prime phosphatase side activity |
| Detection Method | 5 prime hydroxyl termini on single or double stranded DNA and RNA |
| Sample Type | Oligonucleotides; DNA and RNA substrates; NGS adapters |
| Specificity | Inhibited by 7 mM phosphate, pyrophosphate or sodium phosphate at 50% and by 7 mM ammonium sulfate at 75% activity |
| Reaction Conditions / Protocol | Requires Mg2+ and sulfhydryl reagents such as DTT |
| Storage Conditions | -20°C |
| Shelf Life | 1 year |
| Package Specifications | 500 U; 2500 U; 10000 U |
| Key Features | Three activities in one enzyme: 5 prime kinase, 3 prime phosphatase and ADP dependent 5 prime phosphatase exchange |
| Concentration | 10 U/uL |
| Shipping Conditions | Dry ice |
| Readouts / Downstream Assays | Phosphorylation efficiency by ligation assay or end labeling signal |
For in vitro diagnostic use.
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