Product Number: LG06W

Storage

Store at -20℃.

Component

Concentration: 5U/μL

Description

In the presence of Mg²⁺and ATP, T4 DNA ligase can catalyze the formation of phosphodiester bonds between adjacent 5 ‘- phosphate and 3’ – hydroxyl termini on double stranded DNA or RNA. This enzyme can not only catalyze the connection between the flat or viscous ends of double stranded DNA, but also repair single stranded breaks in double stranded DNA, RNA, or DNA/RNA hybrid double stranded DNA, but cannot catalyze the connection between single stranded DNA.

Application

The connection between the sticky and flat ends of double stranded DNA; TA cloning; Cut and repair of double stranded DNA, etc.

Enzyme storage buffer

10 mM Tris-HCl (pH 7.5), 50mM KCl, 1mM DTT, 0.1mM EDTA, 50% (v/v) glycerol.

5 × T4 DNA Ligase Buffer

250 mMTris-HCl (pH 7.5), 50mM MgCl₂, 50mM DTT, 5mM ATP，connection accelerator.

Source

Purified from E. carrying T4 DNA ligase gene Escherichia coli strain.

Inhibitors and thermal inactivation

NaCl or KCl above 200mM can strongly inhibit ligase activity. Heating at 65℃ for 10 minutes or at 70℃ for 5 minutes will deactivate it.

Unit definition

The amount of enzyme required to exchange 1 nmole of 32PPi within 20 minutes at 37℃ is defined as one Weiss unit. The enzyme 1U (Weiss Unit) of this product is equivalent to 200 viscous end units. In T4 DNA ligase reaction buffer, at 16℃ for 30 minutes, the amount of enzyme required to connect 50% of Hind III digested lambda DNA fragments is one viscous terminal unit.

Quality control

The analysis experiment of blue and white spots shows that the number of spots is less than 2%; The detection experiment of excess T4 DNA Ligase mixed supercoiled plasmid showed no nuclease detection; SDS-PAGE detection of recombinant protein purity greater than 99%.