T4 DNA Ligase

0.1 units is defined as the amount of enzyme that is needed to convert 1 pmol (of 18 pmol) of nicked DNA substrate in 20 minutes at 25 °C in a 20 µl reaction volume in a buffer consisting of 62.5 mM Tris-HCl pH 7.5, 10 mM DTT, 1 mM ATP, 0.05 mg/ml BSA and 25 mM KCl.

One Weiss Unit is equivalent to approximately 500 ArcticZymes Units.

56.9 kDa

T4 DNA Ligase can be completely inactivated by incubating at 70°C for 10 minutes.

The enzyme is stable at -20°C for > 1 year in the supplied storage buffer.

The enzyme tolerates at least four freeze-thaw cycles (-80°C) without loss of activity.

10 000 U T4 DNA Ligase was incubated with a supercoiled plasmid (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any transformation of closed circular DNA to nicked DNA.

10 000 U T4 DNA Ligase was incubated with M13 ssDNA (0.5 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of ssDNA degradation.a

10 000 U T4 DNA Ligase was incubated with either 3H-dATP labelled ds or ssDNA (0.5 µg, 500 bp) for 4 hours at 37°C. Acid soluble radioactivity from labelled DNA was not significantly over blank test for either substrate.

5000 U T4 DNA Ligase was incubated with a 2 kb RNA transcript (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of RNA degradation.

5000 U T4 DNA Ligase was analysed in a probe-based qPCR assay detecting the 23S ribosomal subunit in E. coli. No E. coli gDNA could be detected (LOD: < 3 E. coli genomic copies).