E. coli DNA Ligase

• 产品编号：M0205S      品牌：New England Biolabs       原厂货号：M0205S
• 产品分类：克隆 > 限制性内切酶和连接酶 > 连接酶
• 应用分类：

描述：

♦ Specific Activity: 6,000 units/mg

♦ Supplied with 10X Reaction Buffer

Description:
Catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA containing cohesive ends.

Source:
Purified from E. coli strain 594(su^-) carrying the prophage λgt4 lop11 lig⁺Sam 7 (1) by the procedure of Panasenko et al. (2).
 

Applications:

Okayama and Berg cDNA cloning (3)

Reagents Supplied:
E. coli DNA Ligase Reaction Buffer (10X)
 

Heat Inactivation:
65°C for 20 minutes

Specific Activity:
6,000 units/mg

Reaction Conditions: 1X E. coli DNA Ligase Reaction Buffer
Incubate at 16°C.

1X E. coli DNA Ligase Reaction Buffer:
30 mM Tris-HCl
4 mM MgCl₂
26 µM NAD
1 mM Dithiothreitol
50 µg/ml BSA
pH 8.0 @ 25°C

Unit Definition:
One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA (5' DNA termini concentration of 0.12 μM, 300 μg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X E. coli DNA Ligase Reaction Buffer.

Concentration:
10,000 units/ml

Storage Conditions:
10 mM Tris-HCl
50 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

 

原厂资料：

注意事项：

General notes:

Does not ligate RNA to DNA (4).

This enzyme ligates only DNA fragments with cohesive termini.

Usage notes:

Requires NAD⁺ (nicotinamide adenine dinucleotide) as a cofactor, in contrast to other ligases which use rATP.

Ligation of blunt-ended fragments is extremely inefficient. For ligation of blunt-ended fragments use T4 DNA Ligase.