商品编号


EG-1033



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


Ligases




1,000,000 Units

商品信息



Description


T7 DNA Ligase is an ATP-dependent ds DNA ligase from bacteriophage T7. It catalyzes the formation of a phosphodiester bond between a 5′ phosphate and a 3′ hydroxyl termini in duplex DNA. Cohesive end ligation and nick sealing can be efficiently catalyzed by T7 DNA Ligase. However, it is less efficient than T4 and T3 DNA Ligases for blunt end ligationCohesive ligation with T7 DNA Ligase can be completed in less than 10 minutes




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 T7 DNA Ligase gene



Components



T7 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 T7 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 T7 DNA Ligase required to ligate 50% of 100 ng 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.