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T4 DNA Ligase

OverviewTechnical/SpecsLiterature/SupportOrdering Information

for ligation of DNA, TA cloning, and other recombinant DNA applications

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between 5' phosphate and 3' hydroxyl termini in duplex DNA or RNA. This enzyme will ligate DNA from blunt-end and cohesive-end termini (Including sticky ends for TA Cloning) as well as repair single stranded nicks in duplex DNA, RNA, or DNA/RNA hybrids. T4 DNA Ligase is isolated from a recombinant E.coli strain.

Features and Benefits

High Speed: DNA ligations in minutes for cohesive end and in 10 minutes for blunt end DNA at 25°C

Flexibility: Suitable for all common DNA ligations

Reproducibility: Batch manufacturing under strict ISO 9001 quality control system

Enzyme Properties

Heat Inactivation: 70°C for 10 minutes

Application

Blunt or cohesive-end ligation
Repair of nicks in double-stranded nucleic acids

Reagents Supplied

10 x Reaction Buffer: 500 mM Tris-HCI (pH 7.8), 100 mM MgCl₂, 50 mM DTT, 10 mM ATP, 25 μg/ml BSA

Concentration

200 U/μl

Storage Conditions

50% glycerol containing 10 mM Tris-HCI (pH 7.5), 50 mM KCI, 1 mM EDTA, 10 mM 2-Mercaptoethanol

Store Temperature

-20°C

Unit Definition

0.01 Weiss unit of enzyme is defined as the amount of enzyme required to give 90% ligation of Hind III fragments of lambda DNA in 30 minutes, at 16°C in 20 μl of the assay mixture.

Lane 1: DNA fragment (digested with EcoR V)
Lane 2, 3, 4, 5: T4 DNA Ligase 1 U, 16°C, 10, 20, 30 and 60 minutes
Lane 6, 7, 8, 9: T4 DNA Ligase 1 U, 25°C, 10, 20, 30 and 60 minutes
Lane 10, 11, 12,13: T4 DNA Ligase 1 U, 37°C, 10, 20, 30 and 60 minutes
Lane 14: Lambda DNA (digested with Hind III)
Lane 15, 16, 17: T4 DNA Ligase 1 U, 16°C, 10, 20 and 30 minutes
Lane 18, 19, 20: T4 DNA Ligase 1 U, 25°C, 10, 20 and 30 minutes
Lane 21, 22, 23: T4 DNA Ligase 1 U, 37°C, 10, 20 and 30 minutes