TDT；Terminal Transferase；Terminal Deoxynucleotidyl Transferase CAS 9027-67-2
Item No.： E0095
Store at： -20℃
English name：TDT；Terminal Transferase；Terminal Deoxynucleotidyl Transferase
Other names: Terminal deoxynucleotidyl transferase; Thymopentin
CAS NO：9027-67-2
Molecular weight：60000
Grade：BR
Source: E. coli cells, containing the gene encoding bovine thymus terminal deoxynucleoside transferase.
Concentration：20u/ul
Activity definition: refers to the amount of enzyme required to mix 1nmol deoxyribonucleic acid into polynucleotide fragment (adsorbed on DE-81) at 37 ℃ and 60 minutes.
Enzyme activity analysis mixture：66mM Potassium dimethyl arsenate（PH7.2），1mM CoCL2, 0.01%(v/v) Triton X-100, 10uM oligo(dT)10, 1mM dTTP和0.4MBq/ml[3h]-dTTP
Preservation solution components：100mM Potassium acetate（PH6.8)、2mM β- mercaptoethanol，0.01%(v/v) Triton X-100 and 50%(v/v) glycerol.
5×Reaction buffer：1M Potassium dimethyl arsenate、125mM Tris，0.05%(v/v) Triton X-100，5mM CoCL2(PH7.2,25℃)
Inhibitors: Metal chelators, ammonium, chloride, iodide and phosphate ions
Deactivation: add EDTA and heat at 70 ℃ for 10 minutes
Quality assurance: The test shows that there is no contamination of internal and external deoxyribonuclease, ribonuclease and phosphatase.
Note: Because it contains CoCl2, TDT reaction buffer is not compatible with downstream applications. The reaction mixture needs to be purified by column centrifugation, phenol / chloroform extraction and ethanol precipitation to remove CoCl2.
Physicochemical properties: Suspension, recombinant enzyme.（Terminal transferase, TdT） is a template free DNA polymerase that catalyzes the binding of deoxynucleotides to the 3 'hydroxyl end of DNA molecules.Single and double stranded DNA molecules with protruding, concave or smooth ends can be used as substrates for TDT.The general operation is: first, open a single site on the vector and keep it warm together with terminal transferase and a dNTP (such as dATP) to add tail, while the other exogenous DNA fragment to be inserted is added with dNTP (dTTP) by complementary and the same method.Then, the two DNA fragments with complementary single stranded tails are annealed to form a hybrid vector to transform the recipient bacteria. Cracks or incisions in heterozygous carriers can be repaired by whole bacteria.
Purpose: biochemical research. Catalyzing the addition of homopolymers at the 3 'end of DNA;3 'terminal marker of DNA.
