商品编号


EG-1042



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


Ligases




200,000 Units

商品信息



Description

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between the terminal 5′ phosphate and a 3′ hydroxyl groups of duplex DNA or RNA. The enzyme efficiently joins blunt and cohesive ends and repairs single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.

Cohesive ligation with T4 DNA Ligase can be completed in less than 10 min.



Applications



Cloning of restriction fragments

Joining linkers and adapters to blunt-ended DNA

Library construction

TA cloning

Recirculization of linear DNA





Source

Purified from a recombinant
E. coli
strain carrying the T4 DNA Ligase gene



Components



T4 DNA Ligase
, 2,000,000 U/ml in storage buffer: 10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol, pH 7.5 @ 25°C.

4 X T4 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 T4 DNA Ligase required to join 50% of 100 ng of DNA fragments with cohesive termini in 50 ?l 1X T4 DNA Ligase Buffer following a 30 minute incubation 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.