T4 DNA Polymerase CAS 9012-90-2
Item No.： E0091
Store at： -20℃
English name：T4 DNA Polymerase
Molecular weight：104kDa，Monomer
Grade: BR
Source: Escherichia coli containing T4 phage clone gene 43
Concentration：5～10u/ul
Definition of activity: It refers to the amount of enzyme required to add 10nmol deoxyribonucleotide into polynucleotide fragment (adsorbed on DE-81) at 37 ℃ and 30 minutes.
Enzyme activity analysis mixture：67mM Tris-HCL(PH8.8), 1mM DTT, 6.7mM MgCL2, 16.7mM (NH4)2SO4, 0.2mg/ml BSA, 0.033mM:various dNTP,dGTP, 0.4MBq/ML(3H).-dTTP and 0.2mM: Thermal denaturation and Nuclease treated bovine thymus DNA
Save buffer components：20mM Potassium phosphate (PH7.5), 200mM KC1, 20mM DTT和50%(v/v) glycerol
5× Reaction buffer：335mM Tris-HCL(PH8.8，25℃), 33mM MgCL2, 5mM DTT, 84mM (NH4)2SO4
Inhibitor: Metal chelator, nucleotide analogue 2 (p-n-butylanilino) - dATP, N2 - (p-n-burylanilino) - dGTP, SH blocking mixture
Inactivation: Heating at 75 ℃ for 10 minutes
Quality control: The test shows that there is no endodeoxyribonuclease pollution.
Physicochemical properties: Suspension.In the presence of templates and primers, T4 DNA polymerase catalyzes the synthesis of DNA along the 5 '→ 3' direction.The enzyme also has 3 '→ 5' exonuclease activity, which is stronger than DNA polymerase I. Unlike E. coli DNA polymerase I, T4 DNA polymerase does not have 5 '→ 3' exonuclease activity.
Purpose: Biochemical research.3 'protruding end smoothing; 5 'protruding end leveling; Subclones after single strand deletion; Synthesis of the second chain in gene location mutation; DNA probes were labeled by replacement synthesis.
