DNALIG-RO

T4 DNA Ligase

solution, from Escherichia coli(NM 989), suitable for activity assay

• Synonym(s): dna ligase, t4
• UNSPSC Code: 12352204
• NACRES: NA.54

Properties

• biological source: Escherichia coli (NM 989)
• Quality Level: 100
• recombinant: expressed in E. coli
• description: recombinant form of the enzyme from T4 phage
• form: solution
• specific activity: ~3000 units/mg protein
• packaging: pkg of 100 U (10481220001 [1 U/μl]); pkg of 500 U (10716359001 [1 U/μl]); pkg of 500 U (10799009001 [5 U/μl])
• manufacturer/tradename: Roche
• parameter: 4-22 °C optimum reaction temp.
• technique(s): activity assay: suitable
• optimum pH: 7.2-7.8
• storage temp.: −20°C

Description

General description

T4 DNA Ligase catalyzes the formation of phosphodiester bonds between neighboring 3′-hydroxyl- and 5′-phosphate ends in double-stranded DNA. Single-stranded nicks in double-stranded DNA are also closed by T4 DNA Ligase.

Contents
T4 DNA Ligase, supplied with 10x concentrated ligation buffer that includes ATP.

Application

Ligation of sticky- and blunt-ended DNA fragments.[1][2][3]

Biochem/physiol Actions

Heat inactivation: The enzyme can be inactivated by heating (65 °C, 10 minutes).

Preparation Note

Activator: Mg²⁺ (10mM), DTT
Working solution: For dilution of the enzyme Roche recommends using a buffer containing the components of the storage buffer: 20 mM Tris-HCl, 60 mM KCl, 1 mM EDTA, 5 mM dithoerythritol, 50% glycerol (v/v), pH 7.5 (4 °C).

Analysis Note

Quality

Tested for the absence of deoxyribonucleases and exonucleases, according to the current Quality Control procedures.

Other Notes

For life science research only. Not for use in diagnostic procedures.

One unit of T4 DNA ligase is the amount of enzyme activity that converts 1 nmol ³²P from pyrophosphate into Norit-absorbable material in 20 minutes at +37 °C. One unit corresponds to 0.2 U determined in the exonuclease III resistance assay. Dilution buffer: 50 mM Tris-HCl, 10 mM dithioerythritol, bovine serum albumin, 500 μg/ml, pH 7.6 (+25 °C).

Volume Activity: 1 x 10³ U/ml; 5 x 10³ U/ml

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): does not flash
• Flash Point(C): does not flash