PRODUCT DESCRIPTION产品描述
Terminal deoxynucleotidyl transferase (TdT)is a template-independent DNA polymerase that catalyzes the addition ofdeoxynucleotides to the 3′hydroxyl terminus of single or double stranded DNA molecules. Thepresence of 1 mM Co2+stimulates the tailing of the 3′-ends of DNA fragments. Thisconstruct is sold as an N-terminal truncation of the terminal transferase geneattached to an N-terminal fusion tag.
Source of Protein蛋白来源
An E. coli strain that carries the clonedterminal transferase gene from calf thymus.
Supplied in
50 mM KPO4
100 mM NaCl
1.0 mM DTT
0.1 mM EDTA
0.1% Triton X-100
50% Glycerol
pH 7.3 @ 25°C
Supplied With
B0120 (10X Green Buffer)
B0220 (2.5 mM CoCl2)
10X Green Buffer (B0120)
200 mM Tris-Acetate
500 mM Potassium Acetate
100 mM Magnesium Acetate
pH 7.9 @ 25°C
Unit Definition单位定义
1 unit is defined as the amount ofpolymerase required to convert 1 nmol of dTTPs into acid insoluble material in1 hour at 37°C.
PRODUCT INFORMATION产品信息
Terminal deoxynucleotidyl Transferase (TdT)
Part Number P7070L
Concentration 20,000 U/ml
Unit Size 6,000 U
SDS Available on request
PRODUCT SPECIFICATION*产品特性
Storage Temperature -25 to -15°C
Test Specification
Purity (SDS-PAGE) >99%
Specific Activity 27,400 U/mg
SS Exonuclease 200 U <5.0%released
DS Exonuclease 200 U <1.0%released
DS Endonuclease 200 U = No conversion
E.coli DNA Contamination 200 U <10 copies
* For a detailed summary of assayconditions and data, refer to the Quality Controls Analysis section.
USAGE INSTRUCTIONS使用说明
Non-templated addition of dNTPs to 3′termini of DNA:
Reaction Set-Up:
Amount Description Final Concentration
5 μL 10X Green Buffer (B0120) 1X
X μL 10 pmol DNA termini (10-100 ng) 1-10ng/μL
x μL Deoxynucleotide solution 200 μM
1 μL Terminal Transferase (20 U/ μL) 30 U/μL
X μL Type I Water N/A
50 μL Total Volume
Incubate at 37°C for 30 minutes.
Inactivate the TdT and stop the reaction byheating to 70°C for 10minutes.
NOTES
Co2+ increases the nucleotideincorporation efficiency of pyrimidines, and at blunt and 3′ recessed ends. However, theaddition of dNTPs to 3′-overhanging ends is more efficient than with 3′-recessed or blunt ends. TdTrequires a free 3′-hydroxylgroup in order to make a non-templated nucleotide addition.
With limited efficiency, Terminal Transferasewill incorporate ribonucleotides, biotinylated, and dideoxynucleotides in thepresence of Co 2+ .
Terminal Transferase incorporates dATP anddTTP with a 5-fold higher efficiency than dCTP and dGTP, as evidenced by thefollowing Km values for nucleotides:
Base Km
dATP 100μM
dTTP 100μM
dCTP 500μM
dGTP 500μM
REFERENCES参考文献
Deng, G.R. and Wu, R. 1983 Meth. Enzymol.100:96-116.
Limitations of Use适用范围
This product was developed, manufactured,and sold for in vitro use only. The product is not suitable for administrationto humans or animals.
www.hyqbio.com.cn