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商品編號


EDLA-200



品牌


MCLab



公司


MCLAB.Inc



公司分類(lèi)


DNA/RNA Ligases



Concentration

10,000 U/ml


Size

5,000 units

商品信息


E. coli
DNA Ligase is a NAD
+
-dependent enzyme that catalyzes the formation of phosphodiester bonds between complementary 3'-hydroxyl and 5'-phosphoryl termini of dsDNA.




Description:
E. coli
DNA Ligase is a NAD
+
-dependent enzyme that catalyzes the formation of phosphodiester bonds between complementary 3'-hydroxyl and 5'-phosphoryl termini of dsDNA. The enzyme works best with cohesive dsDNA ends and is also active on nicked DNA. Blunt ends can be ligated in the presence of condensing reagents such as polyethylene glycol or Ficoll
?
. A 10x Reaction Buffer is provided with the enzyme.
Source:
E. coli
strain containing an overproducing clone of
E. coli
DNA Ligase.
Application:
- Ligation for cloning
- Okayama and Berg
CDN
A cloning
Unit Definition:
One unit of
E. coli
DNA Ligase is defined as the amount of enzyme required to provide ligation (>50%) of Hind III digested phage lam
BD
a DNA (5'-DNA termini concentration of 0.12 ?M, 300 ?g/ml) in 30 minutes at 16°C under standard assay conditions.
Reaction Conditions:
10x
E. coli
DNA Ligase Reaction Buffer
Incubate at 16°C
10x
E. coli
DNA Ligase Reaction Buffer:
300 mM Tris-HCl
40 mM MgCl
2
260 ?M NAD
10 mM DTT
pH 8.0 @ 25°C


Specific Activity:
?10,000 Unit/
m
g


Recommended Storage Condition:
-20°C
References:
1. Zimmerman, S. B. and Pheifer, B. H. (1983) Proc. Natl. Acad. Sci., USA 80, 5852-5856.
2. Okayama, H. and Berg, P. (1982) Mol. Cell. Biol. 2, 161-170.