Terminal Deoxynucleotidyl Transferase
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.
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.
- Incorporates modified nucleotides (e.g., fluorescein-, biotin-, aminoallyl-labeled nucleotides)
- 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)
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)|
|Definition of Activity Unit||
|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.|
|Source||E.coli cells carrying a cloned gene encoding calf thymus terminal deoxynucleotidyl transferase.|
|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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