R2D Ligase, 5000U, 10 U/uL

ArcticZymes RNA to DNA Ligase (ArcticZymes R2D LigaseTM) is the first ligase on the market that is able to ligate DNA to 5’-phosphorylated ends of RNA in the presence of a DNA template positioning the joinable ends. With its unique substrate specificity, ArcticZymes R2D Ligase allows the development of new technologies in molecular biology research, diagnostics, and manufacturing. Properties Source:  Recombinantly expressed in E. coli. Unit Definition:  One milliunit is defined as the amount of enzyme needed to ligate 1 pmol (of 18 pmol) of a 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 (25°C), 5 mM MgCl2, 1 mM ATP, 10 mM DTT, 0.025 mg/ml BSA and 25 mM KCl. Size: 52.9 k Da Storage and Stability: The enzyme is stable at -20°C for 1 year in the supplied storage buffer. Quality Control ArcticZymes is dedicated to the quality of our products. R2D DNA Ligase is manufactured at our ISO 13485 certified facility in Norway. dsDNA endonuclease activity 2.5 U R2D 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. ssDNA endonuclease activity 2.5 U R2D 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. Exonuclease activity 2.5 U R2D 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. RNase activity 2.5 U R2D 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. E. coli gDNA contamination 2.5 U R2D 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).