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D2886

Sigma-Aldrich

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:
9015-85-4
Enzyme Commission number:
6.5.1.1 (BRENDA, IUBMB)
EC Number:
232-770-0
MDL number:
MFCD00163508
UNSPSC Code:
12352204
NACRES:
NA.53

grade

for molecular biology

form

buffered aqueous glycerol solution

specific activity

4,000 U/mL

mol wt

68 kDa

UniProt accession no.

P00970

storage temp.

−20°C

Gene Information

bacteriophage T4 ... 30(1258680)

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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. 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.

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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Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H334

Precautionary Statements

P261 - P284 - P501

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

Regulatory Listings

Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.

JAN Code

D2886PROC:
D2886-100UN-KC:
D2886-100UN:
D2886-500UN:
D2886-VAR:
D2886-BULK:
D2886-BULK-LBL:
D2886-300UN:
D2886-250UN-KC:
D2886-125UN:
D2886-500UN-KC:

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Sambrook, J., et al.
Molecular Cloning: A Laboratory Manual, 1-1 (1989)
Athena Kantartzis et al.
Cell reports, 2(2), 216-222 (2012-09-04)
Trinucleotide repeat (TNR) expansions are the underlying cause of more than 40 neurodegenerative and neuromuscular diseases, including myotonic dystrophy and Huntington's disease. Although genetic evidence points to errors in DNA replication and/or repair as the cause of these diseases, clear
Adam B Robertson et al.
The Journal of biological chemistry, 287(39), 32953-32966 (2012-08-01)
The Escherichia coli very short patch (VSP) repair pathway corrects thymidine-guanine mismatches that result from spontaneous hydrolytic deamination damage of 5-methyl cytosine. The VSP repair pathway requires the Vsr endonuclease, DNA polymerase I, a DNA ligase, MutS, and MutL to
Justin L Sparks et al.
Molecular cell, 47(6), 980-986 (2012-08-07)
Ribonucleotides are incorporated into DNA by the replicative DNA polymerases at frequencies of about 2 per kb, which makes them by far the most abundant form of potential DNA damage in the cell. Their removal is essential for restoring a
Engler, M.J. and Richardson, C.C. et al.
The Enzymes, 5, 3-3 (1982)
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