85-1360-99　Terminal Deoxynucleotidyl Transferase (TdT) 2500U　T2750

特徴

• TdT is a template-independent DNA polymerase located in the cell nucleus which catalyses irreversible terminal addition, via polymerization, of deoxynucleotides at the 3' hydroxyl ends of oligo or polydeoxynucleotide initiators accompanied by the release of inorganic phosphate.
• Applications:
• Oligo and DNA 3’-terminal labeling
• Homopolymeric tailing of linear duplex DNA with any type of termini
• TUNEL assay (in situ localization of apoptosis)
• 5’-RACE
• Components:
• Terminal Deoxynucleotidyl Transferase: 1x500U or 1x2500U
• Reaction Buffer (5X): 1x0.4ml or 2x1ml1M potassium cacodylate, pH 7.2 (25°C), 5mM CoCl2, 0.5mM DTT, 0.05% Triton X-100
• Storage Buffer:100mM potassium acetate, pH 6.8, 2mM 2-mercaptoethanol, 0.01% Triton X-100, 50% glycerol.
• Unit Definition:1 unit of the enzyme iincorporates 1 nanomole of deoxythymidylate into a polynucleotide fraction in 60 minutes at 37ºC.
• Note: Due to the presence of CoCl2, the TdT Reaction Buffer is incompatible with downstream applications. It is necessary to remove CoCl2 from the reaction mixture by spin column or phenol/chloroform extraction and subsequent ethanol precipitation.
• Quality Control:Tested for absence of exodeoxyribonucleases, endodeoxyribonucleases ribonucleases, and phosphatases.
• Inhibitors:Metal chelators, ammonium chloride, iodide and phosphate ions.
• Inactivators:Inactivated by heating at 70°C for 10 mins. or by addition of EDTA.
• Storage and Stability:Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.