Terminal Deoxynucleotidyl Transferase

Kód produktu: EP0161 Kód výrobce: EP0161 Kód dodavatele: {C36081CB-61AD-4762-8CF7-FA155D7DA74D} Výrobce: UAB Fermentas
2 061,84 Kč
1 704,00 Kč bez DPH
do týdne
500 Units

.

LO certified Recombinant enzyme Thermal inactivation at 70°C in 10 min
Terminal Deoxynucleotidyl Transferase (TdT) catalyzes the template-independent addition of deoxyribonucleotides to the 3'-OH terminus of DNA molecules.

Zobraz detailní popis

Detailní popis

Thermo Scientific Terminal Deoxynucleotidyl Transferase (TdT), a  template-independent DNA polymerase, catalyzes the repetitive addition of deoxyribonucleotides to the 3'-OH of oligodeoxyribonucleotides and single-stranded and double-stranded DNA (see Reference 1). TdT requires an oligonucleotide of at least three nucleotides to serve as a  primer. With RNA as template TdT shows variable performance which strongly depends upon the tertiary structure of acceptor RNA 3'-end and the nature of nucleotide. Generally, it is lower than using DNA as a  template.

Highlights

  • Incorporates modified nucleotides (e.g., fluorescein-, biotin-, aminoallyl-labeled nucleotides)

Applications

  • Production of synthetic homo- and heteropolymers (see​ Reference 1)
  • Homopolymeric tailing of linear duplex DNA with any type of 3'-OH terminus (see​ References 2, 3)
  • Oligodeoxyribonucleotide and DNA labeling (see​ References 2, 4-8)
  • 5'-RACE (Rapid Amplification of cDNA Ends) (see​ Reference 9)
  • In situ localization of apoptosis (see​ Reference 10)

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.

5X Reaction Buffer 1 M potassium cacodylate, 125 mM Tris, 0.05% (v/v) Triton X-100, 5 mM CoCl2 (pH 7.2 at 25°C)
CategoryName
Concentration 20 U/µL
Definition of Activity Unit
  • One unit of the enzyme catalyzes the incorporation of 1 nmol of deoxythymidylate into a polynucleotide fraction (adsorbed on DE-81) in 60 min at 37°C.
  • Enzyme activity is assayed in the following mixture: 200 mM potassium cacodylate (pH 7.2), 1 mM CoCl2, 0.01% (v/v) Triton X-100, 10 µM oligo(dT)10, 1 mM dTTP, and 0.4 MBq/mL [3H]-dTTP.
Hazardous No
Hazardous: No
Inactivation Inactivated by heating at 70°C for 10 min or by addition of EDTA.
Inhibition Inhibitors: metal chelators, ammonium, chloride, iodide, phosphate ions
Quality Control The absence of endo-, exodeoxyribonucleases, phosphatases and ribonucleases confirmed by appropriate quality tests.
Shelf Life:
Shipping Condition:
Shipping Information
Source E.coli cells carrying a cloned gene encoding calf thymus terminal deoxynucleotidyl transferase.
SpecificationName
SpecificationValue
Storage Buffer The enzyme is supplied in 100 mM potassium acetate (pH 6.8), 2 mM 2-mercaptoethanol, 0.01% (v/v) Triton X-100 and 50% (v/v) glycerol.
Storage Condition -20 C
Storage Condition: -20 C

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