New England Biolabs, R0644S, Nt.BspQI

Related Categories Nicking Endonucleases,, Restriction Endonucleases: N-O Specification Unit Definition One unit is defined as the amount of enzyme required to convert 1 μg of supercoiled pUC19 DNA to open circular form in 1 hour at 50°C in a total reaction volume of 50 μl. Reaction Conditions 1X NEBuffer™ r3.1 Incubate at 50°C 1X NEBuffer™ r3.1 100 mM NaCl 50 mM Tris-HCl 10 mM MgCl2 100 µg/ml Recombinant Albumin (pH 7.9 @ 25°C) Activity in NEBuffers NEBuffer™ r1.1: <10% NEBuffer™ r2.1: 25% NEBuffer™ r3.1: 100% rCutSmart™ Buffer: 10% Diluent Compatibility Diluent B Storage Buffer 10 mM Tris-HCl 300 mM NaCl 1 mM DTT 0.1 mM EDTA 500 µg/ml BSA 50% Glycerol pH 7.4 @ 25°C Heat Inactivation 80°C for 20 minutes Methylation Sensitivity dam methylation: Not Sensitive dcm methylation: Not Sensitive CpG Methylation: Not Sensitive Activity at Temperature @37°C: 50% FAQ Q: Are there any nicking enzymes that are used on the Irys® system from BioNano Genomics®? A: Nb.BssSI and Nt.BspQI are nicking endonucleases that cleave only one strand of DNA on a double-stranded DNA substrate. They can be used for genome mapping, assembly and SV detection on the Irys system from BioNano Genomics. For more information please visit www.BioNanogenomics.com or contact BioNano at : support@bionanogenomics.com Q: Is this enzyme sensitive to dam, dcm or mammalian CpG methylation? A: No. This enzyme is not sensitive to dam, dcm, or mammalian CpG methylation. For up-to-date information about methylation sensitivities, please visit Dam-Dcm and CpG Methylation or REBASE. Q: Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers? A: NEB is excited to announce that we have switched BSA-containing reaction buffers (NEBuffer 1.1, 2.1, 3.1 and CutSmart® Buffer) to Recombinant Albumin-containing buffers (NEBuffer r1.1, r2.1, r3.1 and rCutSmart™ Buffer). We are also in the process of moving all restriction enzyme formulations to contain rAlbumin. We feel that moving away from animal-containing products is a step in the right direction and are able to offer this enhancement at the same price.