Product Introduction
T4 Polynucleotide Kinase (T4 PNK) catalyzes the transfer and exchange of Pi from the γ position of ATP to the 5'-hydroxyl terminus of polynucleotides (double-stranded and single-stranded DNA and RNA) and nucleoside 3'-monophosphate. T4 polynucleotide kinase also catalyzes the removal of 3′-phosphoryl groups from 3′-phosphate-based polynucleotides, deoxynucleoside 3′-monophosphate, and deoxynucleoside 3′ -diphosphate. When ADP is present, T4 PNK has 5'-end phosphatase activity, catalyzing the exchange of 5'-P-oligomeric/polymeric nucleotides and ATP terminal 5'-phosphate groups. It is suitable for constructing DNA libraries and producing probes with terminal labeling
Application
1. End labeling DNA or RNA for probes and DNA sequencing;
2. 5' phosphorylation of DNA/RNA for subsequent ligation;
3. Phosphorylate the 5´ end of the 3´ phosphorylated mononucleotide to prepare the pNp substrate for addition to the 3´ end of DNA or RNA;
4. Label the 5´ end of oligonucleotides with a phosphate group at the 3´ end.
Features
Phosphorylation of the 5′ end of DNA or RNA for ligation reactions
Terminal labeling of DNA or RNA for use as probes and DNA sequencing
Components
Component | K013-A (50U) | K013-B (2500U) | K013-C (10000U) |
T4 Polynucleotide Kinase (10U/µL) | 50 µL | 250 µL | 1 mL |
10X T4 PNK Buffer | 50 µL | 1 mL | 1 mL x 2 |
Storage Condition
All reagents should be stored at -20°C.