English name: T4 DNA Polymerase
CAS number: 9012-90-2
Molecular weight: 104kDa, single
Source: T4 phage clone 43 gene of Escherichia coli
Activity definition: refers to 37 degrees, 30 minutes 10nmol Deoxyribose Nucleic acid polynucleotide fragments mixed (adsorption on DE-81) the amount of enzymes
Enzyme:67mMTris-HCL(PH8.8),1mMDTT,6.7mMMgCL2,16.7mM(NH4)2SO4,0.2mg/mlBSA,0.033mM activity analysis of mixture of dNTP, dGTP, 0.4MBq/ML (3H).-dTTP and 0.2mM thermal denaturation of nucleic acid enzymes and bovine thymus DNA
Save buffer components: 20mM potassium phosphate (PH7.5), 200mM KC1 20mM DTT and 50% (v/v) glycerol
5* reaction buffer: 335mM Tris-HCL (PH8.825), 33mM, MgCL2, 5mM DTT, 84mM (NH4) 2SO4
Inhibitors: metal chelator, 2 nucleotide analogues (p-n-butylanilino)-dATP, N2-(p-n-burylanilino)-dGTP, SH-mixture
Deactivation: 75 deg c for 10 minutes
Quality control: the enzyme tests showed no DNA contamination
Traits: suspension. Recombinant enzymes and conditions existing in the template and primers, T4 DNA polymerase-catalyzed synthesis of DNA along the 5' → 3'. The enzyme also has the sequense of exonuclease activity, the activity of DNA polymerase I is strong. T4 DNA polymerase is not like Escherichia coli DNA polymerase I, do not have 5' → 3-exonuclease activity.
Use: biochemical studies. 3' highlights smoothing at the end; 5' prominent end filling; Deleted and subcloning of single chain; Gene mutation in the the second strand synthesis; Synthesis by replacement (replacement synthesis) to label DNA probes.