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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100 UNITS
PLN 478.00
500 UNITS
PLN 1,710.00
PLN 478.00
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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
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grade
Molecular Biology
form
buffered aqueous glycerol solution
specific activity
4,000 U/mL
mol wt
68 kDa
UniProt accession no.
storage temp.
−20°C
Quality Level
Gene Information
bacteriophage T4 ... 30(1258680)
Related Categories
1 of 4
This Item | |||
|---|---|---|---|
| specific activity 4,000 U/mL | specific activity - | specific activity - | specific activity - |
| grade for molecular biology | grade - | grade for molecular biology | grade for molecular biology |
| Gene Information bacteriophage T4 ... 30(1258680) | Gene Information bacteriophage T4 ... B-GT(1258765) | Gene Information - | Gene Information Escherichia coli K12 ... polA(948356) |
| form buffered aqueous glycerol solution | form aqueous solution | form buffered aqueous glycerol solution | form buffered aqueous glycerol solution |
| mol wt 68 kDa | mol wt 41.6 kDa | mol wt 33 kDa | mol wt 103 kDa |
| UniProt accession no. | UniProt accession no. | UniProt accession no. - | UniProt accession no. |
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]
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.
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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M J Moore et al.
Science (New York, N.Y.), 256(5059), 992-997 (1992-05-15)
A simple and efficient method for synthesizing long, site-specifically modified RNA molecules was developed whereby segments of RNA were joined with the use of bacteriophage T4 DNA ligase. A single hydrogen or O-methyl group was substituted for the 2'-hydroxyl group
Xin Cheng et al.
European journal of cell biology, 91(10), 782-788 (2012-08-04)
Translocation of mitochondrial DNA (mtDNA) fragments to the nucleus and insertion of those fragments into nuclear DNA has been observed in several organisms ranging from yeast to plants and mammals. Disruption of specific nuclear genes by de novo insertions of
J R Rusche et al.
Nucleic acids research, 13(6), 1997-2008 (1985-03-25)
Hexamine cobalt chloride (HCC) increases the efficiency of blunt end ligation by T4 DNA ligase about 50 fold. Maximum stimulation occurs when standard buffers for ligation are supplemented with 1 mM HCC. All the ligation events are intermolecular regardless of
Wenxin Gu et al.
Bioorganic & medicinal chemistry letters, 22(11), 3693-3698 (2012-05-09)
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
Engler, M.J. and Richardson, C.C. et al.
The Enzymes, 5, 3-3 (1982)
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