Not available outside of the UK & Ireland.

Application

Ligation of sticky- and blunt-ended DNA fragments.

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.ContentsT4 DNA Ligase, supplied with 10x concentrated ligation buffer that includes ATP.

Other Notes

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

Preparation Note

Activator: Mg²⁺ (10mM), DTTWorking 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).

Quality

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

Specificity

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

Unit Definition

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