T4 DNA Ligase

Product Overview

Description

T4 DNA Ligase

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA with blunt or cohesive end. The enzyme repairs single strand nicks in duplex DNA, RNA or DNA/RNA hybrids. It has no activity on single-stranded nucleic acids. T4 DNA Ligase requires ATP as cofactor.

 

 

Cat. No.	FL101
	FL101-01	10000 units
	FL101-02	2×10000 units
Storage	at -20°C for two years
Shipping	Dry ice (-70 ℃)
Concentration:	200 units/μl
Unit Definition	One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of λDNA (5´ DNA termini concentration of 0.12 μM, 200 μg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1×T4 DNA Ligase Reaction Buffer.
Source	E.coli strain carrying T4 DNA ligase gene
Quality Control	SDS-PAGE test: purity >90%, functional absence of endonucleases and exonucleases activities
Applications	• Cloning blunt end or cohesive end fragments
	• Ligating linkers or adapters to blunt end DNA
Storage Buffer	10 mM Tris-Cl (pH 7.4), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol
Components	T4 DNA Ligase (200 units/μl), 5×T4 DNA Ligase Buffer [250 mM Tris-Cl (pH 7.5), 50 mM MgCl2, 5 mM DTT, 5 mM ATP, 125 μg/ml BSA, Enhancer ]
Notes	• Suggested molar ratio of insert DNA to vector DNA is at 3:1-10:1.
	• Mix 5×T4 DNA Ligase Buffer completely before use.
	• T4 DNA Ligase is unstable for keeping on ice for a long time. It is suggested taking out when needed for use and place back immediately after use.

 

Reaction System 

Vector DNA	X μl
Insert DNA	X μl
5×T4 DNA Ligase buffer	2 μl
T4 DNA Ligase	0.5-1 μl
ddH2O	to 10 μl

 

 

• Cohesive ends ligation: incubate at 25^oC for 10 minutes.

• Blunt ends ligation: incubate at 25^oC for 2 hours, or overnight at 16^oC.

• Cohesive and blunt ends ligation: incubate 25^oC for 2 hours.

 

 

 

 

 

 

Packaging