Shanghai Yiji Industrial Co., Ltd. is a high-tech enterprise integrating R & D, sales, and technical services. It sells and produces a variety of medical research reagents, mainly engaged in ELISA kits, cells, serum, antibodies, biological reagents, culture media , And other products, please call for details.
English name: TDT; Terminal Transferase; Terminal Deoxynucleotidyl Transferase
Other names: terminal deoxynucleotide transferase
CAS number: 9027-67-2
Molecular weight: 60000
Level: BR
Source: E. coli cells, containing the gene encoding the end deoxynucleoside transferase of bovine thymus Concentration: 20u / ul
Activity definition: refers to the amount of enzyme required to incorporate 1 nmol of deoxyribonucleic acid into a polynucleotide fragment (adsorbed on DE-81) at 37 ° C for 60 minutes. Enzyme activity analysis mixture: 66 mM potassium cacodylate ( PH7.2), 1mM CoCL2, 0.01% (v / v) Triton X-100, 10uM oligo (dT) 10, 1mM dTTP and 0.4MBq / ml [3h] -dTTP
Storage solution components: 100 mM potassium acetate (PH6.8), 2 mM β-mercaptoethanol, 0.01% (v / v) Triton X-100 and 50% (v / v) glycerol
5 × reaction buffer: 1M potassium cacodylate, 125mM Tris, 0.05% (v / v) Triton X-100, 5mM CoCL2 (PH7.2, 25 ℃)
Inhibitor: metal chelating agent, ammonium, chloride, iodide and phosphate ion inactivation: adding EDTA, heating at 70 ℃ for 10 minutes Quality assurance: test shows no endonuclear and exonuclease DNA, ribonuclease, phosphate Enzyme contamination Note: Due to the presence of CoCL2, the TdT reaction buffer is not compatible with downstream applications. Column centrifugation or phenol / chloroform extraction and ethanol precipitation should be used to purify the reaction mixture to remove CoCL2
Properties: suspension, recombinant enzyme. Terminal transferase (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, incubate it with the terminal transferase and a dNTP (such as dATP) to add tail, and another foreign DNA fragment to be inserted is used with complementary Adding dNTP (dTTP) with the same method. Next, the above two DNA fragments with complementary single-stranded tails are annealed to each other to form a hybrid vector to transform the recipient bacteria. The cracks or cuts in the hybrid carrier can be repaired by the whole bacteria.
Uses: biochemical research. Catalyze the addition of homopolymers at the 3 ends of DNA; label the DNA at the 3 ends and store: -20 ° C
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