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D2886 Sigma

DNA Ligase from T4-infected Escherichia coli

buffered aqueous glycerol solution

Synonym: Polydeoxyribonucleotide Synthase, Polynucleotide Ligase, T4 DNA Ligase

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Properties

Related Categories 6.5.x.x Phosphoric ester, 6.x.x.x Ligases, Application Index, Biochemicals and Reagents, Core Bioreagents,
grade   for molecular biology
form   buffered aqueous glycerol solution
activity   4,000 U/mL
mol wt   mol wt 68 kDa
storage temp.   −20°C
Gene Information   bacteriophage T4 ... 30(1258680)

Description

Components

T4 DNA Ligase is supplied in a solution containing 10 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM DTT, and 50% (v/v) glycerol.

Application

Suitable for:
• Ligation of blunt ended or cohesive DNA fragments
• Ligation of cloning vector and restriction insert fragments
• Seal nicks in double stranded DNA and RNA or DNA/RNA hybrids
• Couple RNA single strands by bridging oligonucleotide adapters

Other Notes

T4 DNA Ligase is inactivated by heating at 65 °C for 10 minutes.

Unit Definition

One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmole of P32 from pyrophosphate into ATP as Norit-absorbable material in 20 minutes at 37°C.

Biochem/physiol Actions

T4 DNA Ligase forms an energy dependent phosphodiester linkage between the termini of adjacent polynucleotides of duplex DNA. The ligation reaction requires ATP as a cofactor. Ligation of blunt-ended fragments requires higher enzyme concentration and can be facilitated by using PEG in the reaction mixture. The enzyme requires a 3′ hydroxyl and 5′ phosphate for ligation. Self-ligation of vector DNA can be prevented by dephosphorylation with alkaline phosphatase. T4 ligase plays an active role in repair of DNA and RNA nicks.

Price and Availability

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3
Protocols & Articles

Related Content

Enzymes & Proteins

Application Index | Enzyme Index | Substrate Index | Inhibitor Index | Cofactor Index | Lectin Index
Keywords: Cell signaling, Diagnostic, Drug discovery, Molecular biology

Peer-Reviewed Papers
15

References

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