Polynucleotide kinase

Product Information
Stock solution: 10 mM Tris-HCl, 50 mM KCl, Glycerol, 0.1 μM ATP, pH 7.4
Reaction solution: 700 mM Tris-HCl, 100 mM MgCl₂, 50 mM DTT
Instructions
1) Radioactive phosphorylation labeling is performed using T4 PNK to establish the following reaction system:
DNA up to 50 pmol 5' -termini, Reaction Buffer 5 μL, ATP (10 mM) 50 pmol [y-32P]ATP, T4 polynucleotide kinase 2 μL (20 U), Nuclease-free Water replenished to 50 μL; Incubate at 37°C for 30 minutes; Heat inactivate at 65°C for 20 minutes.
2) Non-radioactive phosphorylation labeling is performed using T4 PNK to establish the following reaction system:
DNA up to 300 pmol 5' -termini, Reaction Buffer 5 μL, ATP (10 mM) 5 μL, T4 polynucleotide kinase 1 μL (10 U), Nuclease-free Water replenished to 50 μL; Incubate at 37°C for 30 minutes; Heat inactivate at 65°C for 20 minutes.
Notes
1) After receiving this product, please try to avoid repeated freezing and thawing.
2) The Reaction Buffer does not contain ATP and needs to be added by yourself.
3) CTP, GTP, TTP, UTP, dATP and dTTP can all replace ATP as phosphoric acid donors.
4) Adding ADP with a final concentration of 100 μM to the reaction system can still maintain a sufficient incorporation level.
5) When using the enzyme, it is advisable to keep it on ice. After use, it should be immediately returned to -20°C.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.