商品编号


EG-1043



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


Ligases




1,000,000 Units

商品信息



Description


T3 DNA Ligase is an ATP-dependent ds DNA ligase from bacteriophage T3. It catalyzes the formation of a phosphodiester bond between a 5′ phosphate and a 3′ hydroxyl termini in duplex DNA. T3 DNA Ligase can efficiently catalyze ligation of both cohesive ends and blunt ends and nick sealing.

T3 DNA Ligase is 5-fold more efficient than T4 DNA ligase for ligation of cohesive DNA fragments, but not as active as T4 DNA ligase for blunt-ended DNA fragments, although the blunt end ligation can be enhanced by the addition of 15% PEG 6000 to the reaction. T3 DNA Ligase displays a higher efficiency for joining A/T overhangs than C/G matched ends, therefore it can be used for TA cloning. Cohesive ligation with T3 DNA Ligase can be completed in less than 10 min.




Applications



Cloning of DNA fragments generated by restriction enzyme digestion

Adding linkers or adapters to dsDNA

Circularization of linear DNA

Nick-sealing in dsDNA

Site-directed mutagenesis

Golden Gate assembly of DNA fragments such as Transcription Activator-Like Effector Nucleases (TALEN)





Source

Purified from a recombinant
E. coli
strain carrying the T3 DNA Ligase gene



Components



T3 DNA Ligase:
3,000,000 U/ml in storage buffer: 20 mM Tris-HCl, 50 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol, pH 7.5 @ 25°C .

4X T3 DNA Ligation Buffer
: 250 mM Tris-HCI, 40 mM MgCl
2
, 4 mM DTT, 4 mM ATP, 30% PEG 6000, pH 7.6 @ 25°C





Unit Definition

1 unit is defined as the amount of T3 DNA Ligase required to ligate 50% of 100 ng HindIII DNA fragments with cohesive termini in 30 minutes at 23°C



Quality Control



The absence of endo-, exodeoxyribonucleases, phosphatases, and ribonucleases confirmed by appropriate quality tests.

Functionally tested for the capacity to join cohesive- and blunt-end DNA fragments.





Storage

-20°C.