Terminal Deoxynucleotidyl Transferase, 50 μL

Product Information

Product Description/Introduction

This product Terminal Deoxynucleotidyl Transferase, abbreviated as TdT, is a template independent DNA polymerase that can catalyze the addition of dNTP to the 3 'hydroxyl end of oligonucleotides, single stranded or double stranded DNA. It can catalyze oligonucleotides with a minimum length of 3 nucleotides. TdT can also add NTP to the 3 'hydroxyl end of RNA, but its catalytic activity towards RNA templates is weaker than that towards DNA templates. This product is mainly used in our company's Tunel Cell Apoptosis Detection Kit (item number: G1501、G1502、G1504、G1505、G1507）。 The TdT produced by our company is obtained through evolutionary screening and has higher enzyme activity and a wider substrate spectrum compared to the wild type.

Applications: synthesis of homopolymers and heteropolymers; Linear double stranded DNA with 3 'hydroxy end homopolymer and tail; Oligonucleotide or DNA 3hydroxyl terminal labeling; 5'-RACE(rapid amplification of cDNA ends); In situ localization of cell apoptosis.

Source: Recombinant expression of Escherichia coli strain carrying calf thymus gene.

Enzyme activity definition: The amount of enzyme required to incorporate 1 nmol deoxyribonucleotide into the 3 'end of a polynucleotide in a matching reaction buffer at 37 for 1 hour is defined as one enzyme activity unit.

Inactivation or inhibition: Heating at 70 for 10 minutes or adding an appropriate amount of EDTA can inactivate it. Metal ion chelating agents, high concentrations of ammonium ions, chloride ions, iodide ions, phosphate ions, heparin, etc. can all inhibit its activity.

Purity and concentration: Purity 95% by SDS-PAGE; no DNase RNase； Endogenous nucleic acid residue<1 pg/μL (qPCR detection); 20 U/μL.

Enzyme storage buffer: 60 mM Potassium phosphate, 150 mM KCl, 1 mM DTT, 0.5% Triton X-100, 50% Glycerol, pH 7.2.

Storage and Shipping Conditions

Ship with wet ice; store at -20°C, valid for 12 months.

Product Content

Assay Protocol / Procedures

1, This TdT can be directly used for our company's Tunel Cell Apoptosis Detection Kit (item number: G1501、G1502、G1504、G1505、G1507）； It can also be used for DNA end labeling and tail addition reactions.Set up the following reaction.

2, DNA 3'end marker:

After setting up the reaction system according to the table above, gently mix the centrifuged precipitate liquid and incubate the reaction system at 37 for 20 minutes. After the reaction is complete, incubate the system at 70 for 20 minutes to terminate the reaction.(The efficiency of DNA 3' end labeling is related to the type of 3' end, and the labeling efficiency of the 3 'protruding end is significantly higher than that of the 3' indented or flat end)

3, Tail addition at the end of DNA:

After setting up the reaction system according to the table above, gently mix the centrifuged precipitate liquid and incubate the reaction system at 37 for 25 minutes. After the reaction is complete, incubate the system at 70 for 20 minutes to terminate the reaction. (According to the above reaction conditions, approximately 110 dA or dT can be added to the 3 'hydroxyl end of each DNA, or approximately 25 dC or dG can be added)

Note

1. When using enzyme products, they should be placed on ice. After use, they should be immediately stored at -20 , and it is recommended to store them separately.

2. Due to the presence of CoCl2 in the 5×Reaction Buffer, it is not compatible with downstream applications. The reaction mixture needs to be purified using column centrifugation or phenol/chloroform extraction and ethanol precipitation methods to remove CoCl2.

3. For your safety and health, please wear a lab coat and disposable gloves during the operation.

For Research Use Only!

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