PRODUCT DESCRIPTION产品描述
T3 DNA Ligase catalyzes the formation of aphosphodiester bond between a 5′ phosphate and a 3′ hydroxyl termini in duplex DNA. The enzyme will join blunt end andcohesive end termini as well as repair single stranded nicks in duplex DNA. Inthe absence of 20-30% PEG 6000, T3 DNA Ligase displays a very low efficiencyfor blunt-ended ligation. (1) T3 DNA Ligase displays a higher efficiency forjoining A/T overhangs than C/G matched ends. (1) T3 DNA Ligase retains 95% ofits activity in 1.0 M NaCl or KCl, with an optimal concentration of 300 mM(1).
Source of Protein蛋白来源
A recombinant E. coli strain carrying theT3 DNA Ligase gene.
Supplied in
20 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
50% glycerol
pH 7.5 @ 25°C
Supplied With:
B1010 (2X Rapid Ligation Buffer)
2X Rapid Ligation Buffer (B1010):
132mM Tris-HCI
20 mM MgCl2
2 mM DTT
2 mM ATP
15% PEG 6000
pH 7.6 @ 25°C
Unit Definition单位定义
1 unit is defined as the amount of T3 DNALigase required to ligate 50% of 100 ng DNA fragments with cohesive termini in30 minutes at 23°C.
PRODUCT INFORMATION产品信息
T3 DNA Ligase
Part Number L6010L,L6010F
Concentration 3,000,000 U/ml
Unit Size 900,000 U, 140,000U
SDS Available on request
PRODUCT SPECIFICATION*产品特性
Storage Temperature -25 to -15°C
Test Specification
Purity (SDS-PAGE) >99%
Specific Activity 3,000,000 U/mg
SS Exonuclease 30,000 U <1.0% released
DS Exonuclease 30,000 U <1.0% released
DS Endonuclease 30,000 U = No conversion
E.coli DNA Contamination 30,000 U <10 copies
* For a detailed summary of assayconditions and data, refer to the Quality Controls Analysis section.
NOTES
T3 DNA Ligase is supplied with 2X RapidLigation Buffer for use in ligation reactions.
The enzyme has also been characterized inthe following buffer, with a 17-fold decrease in specific activity due to theabsence of the crowding agent (PEG 6000):
1X T3 DNA Ligase Buffer:
50 mM Tris-HCl
300 mM NaCl
0.5 mM ATP
1 mM DTT
pH 8.0 @ 25°C
The following information was taken fromthe Cai reference (cited below), which describes the characterization of the T3DNA Ligase:
Optimal pH: 8.0
Optimal reaction temperature: 20°C
Optimal reaction time: >30 minutes(without PEG 6000)
Optimal ionic strength: 300 mM NaCl
Optimal divalent cation/concentration: 2 mMMg++
Optimal cofactor: ATP (0.5 mM)
REFERENCES参考文献
Cai, Liang, et al. (2004) J. Biochem. 135,397-403
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.
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