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Product Introduction
Terminal transferase (TdT) is a template-independent DNA polymerase that catalyzes deoxynucleotide binding to the 3 'hydroxyl end of the DNA molecule. Single-double-stranded DNA molecules with protruding, concave or smooth ends can be used as substrates for TdT, and the tail length can reach 5~300nt. The enzyme is screened by electron heavy framing technology, so that it can react at 45℃, so as to avoid the decrease of tail addition efficiency caused by DNA secondary structure.
The enzyme has been widely used in adding homopolymers to the 3 'end of DNA, labeling DNA3' end with modified bases (such as ddNTP, DIGdUTP), and TDT-mediated dUTP notch end labeling (in situ detection of apoptosis).
Application
1. Oligonucleotide or DNA 3 'hydroxyl terminal labeling
2.DNA tailing
3.5 '- RACE
4. Synthesis of oligomer chains of the same deoxynucleotide
Precautions
1. The activity of TerminalTransferase can be lost with appropriate amount of EDTA.
2. Metal ion chelating agent, relatively high concentration of ammonium ion, chloride ion, iodine ion and phosphate ion all have inhibitory effect on TerminalTransferase activity.
3, the calculation of the number of moles at the end of DNA, the length of 100bp DNA: 1pmol end DNA=0.33ng.
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