Product D2886 - DNA Ligase from T4-infected Escherichia coli is supplied in 20 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM DTT, and 50% (v/v) glycerol.
Unfortuntely, the reaction buffer is not offered. This enzyme cuts in many buffers including buffers for restriction enzymes. In order to work the enzyme needs 1mM ATP added to the ligation reaction.
A recipe for a standard T4 ligation buffer is as follows:
50 mM Tris-HCl, pH 7.8 (T2569)
10 mM MgCl2 (M1028)
10 mM dithiothreitol (D9779)
1 mM ATP (A7699)
25 ug/ml bovine serum albumin (B4287)
D2886
DNA Ligase from T4-infected Escherichia coli
buffered aqueous glycerol solution
Synonym(s):
Polydeoxyribonucleotide Synthase, Polynucleotide Ligase, T4 DNA Ligase
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About This Item
Pricing and availability is not currently available.
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grade
for molecular biology
Quality Level
form
buffered aqueous glycerol solution
specific activity
4,000 U/mL
mol wt
68 kDa
UniProt accession no.
storage temp.
−20°C
Gene Information
bacteriophage T4 ... 30(1258680)
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Related Categories
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
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.[1] Ligation of blunt-ended fragments requires higher enzyme concentration and can be facilitated by using PEG in the reaction mixture.[2] 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.[3]
Components
T4 DNA Ligase is supplied in a solution containing 20 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM DTT, and 50% (v/v) glycerol.
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.
Other Notes
T4 DNA Ligase is inactivated by heating at 65 °C for 10 minutes.
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Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Resp. Sens. 1
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
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A series of 2,6-disubstituted aminoalkoxypyrimidine carboxamides (AAPCs) with potent inhibition of bacterial NAD(+)-dependent DNA ligase was discovered through the use of structure-guided design. Two subsites in the NAD(+)-binding pocket were explored to modulate enzyme inhibitory potency: a hydrophobic selectivity region
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