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
CAS Number:
UNSPSC Code:
12352204
NACRES:
NA.53
EC Number:
232-770-0
MDL number:
Specific activity:
4,000 U/mL
Pricing and availability is not currently available.
grade
Molecular Biology
Quality Segment
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)
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.
Other Notes
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.
T4 DNA Ligase is inactivated by heating at 65 °C for 10 minutes.
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.
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This Item | |||
|---|---|---|---|
| grade Molecular Biology | grade - | grade Molecular Biology | grade Molecular Biology |
| Gene Information bacteriophage T4 ... 30(1258680) | Gene Information bacteriophage T4 ... B-GT(1258765) | Gene Information - | Gene Information Escherichia coli K12 ... polA(948356) |
| specific activity 4,000 U/mL | specific activity - | specific activity - | specific activity - |
| form buffered aqueous glycerol solution | form aqueous solution | form buffered aqueous glycerol solution | form buffered aqueous glycerol solution |
| UniProt accession no. | UniProt accession no. | UniProt accession no. - | UniProt accession no. |
| storage temp. −20°C | storage temp. −70°C | storage temp. −20°C | storage temp. −20°C |
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
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