Products
Recombinant Protein & Enzyme
Modification Enzyme
Phosphatase/Kinase
T4 Polynucleotide Kinase, 500 U
Attribute:
SKU : G3458-500U
Categories : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
T4 Polynucleotide Kinase | G3458-500U | 500 U |
Product Introduction
Product Description: | Servicebio® T4 Polynucleotide Kinase (T4 PNK) catalyzes 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 reaction is reversible. Polynucleotide Kinase also catalyzes the removal of 3´-phosphoryl groups from 3´-phosphoryl polynucleotides, deoxynucleoside 3´-monophosphates and deoxynucleoside 3´-diphosphates |
Applications: | · 5' phosphorylation of DNA/RNA for subsequent ligation. · End labeling DNA or RNA for probes and DNA sequencing. l Removal of 3' phosphoryl groups |
Source: | A E. coli strain that carries the cloned T4 Polynucleotide Kinase gene |
Purity: | 95% by SDS-PAGE |
Endogenous nucleic acid | ＜1 pg/μL by qPCR |
Concentration: | 10 U/μL |
Definition of Activity Unit: | One unit is defined as the amount of enzyme catalyzing the incorporation of 1 nmol of γ-phosphate from ATP to acid-insoluble precipitate in 30 min at 37°C. |
10× Reaction Buffer: | 500 mM Tris-HCl，100 mM MgCl2，5 mM DTT，pH 7.6. |
Storage (Dilution) Buffer: | 10 mM Tris-HCl，50 mM KCl，1 mM DTT，0.1 mM EDTA，50% Glycerol，pH 7.4. |
Inactivation or inhibition | l Inhibitors: metal chelators, phosphate and ammonium ions, KCl and NaCl at a concentration higher than 50 mM. l Inactivated by heating at 65°C for 20 min or by addition of EDTA. |
Storage Conditions: | Store at 20°C up to 12 months. |
Product Contents
Component Number | Component | G3458-500U |
G3458-1 | T4 Polynucleotide Kinase | 50 μL |
G3458-2 | 10х Reaction buffer | 500 μL |
Manual | One copy | |
Assay Protocol / Procedures
Protocol for DNA/RNA 5'-end labeling by T4 PNK.
l Thaw frozen reagents, mix and centrifuge briefly.
l Keep enzymes on ice.
l Keep the 10х Reaction buffer at room temperature.
1. Prepare the following reaction mixture containing:
Component | Volume |
Linear ds DNA or Oligonucleotide | 200 pmol |
10х Reaction buffer | 5 μL |
ATP（10 mM） | 5 μL |
T4 Polynucleotide Kinase | 1 μL |
Nuclease-Free Water | To 50 μL |
Total volume | 50 μL |
2. Mix thoroughly, spin briefly and incubate at 37°C for 30 min.
3. Heat at 65 °C for 20 min.
Note
1. As the T4 DNA ligase buffer contains 1 mM ATP, the T4 Polynucleotide Kinase can share the buffer with T4 DNA ligase.
1. The T4 Polynucleotide Kinase should be shipped on ice when handling, and should be stored at -20 immediately after use.
1. For your safety and health, please wear safety glasses, gloves, or protective clothing.
For Research Use Only!
