New England Biolabs, R0648L, Nb.BsrDI

Note – Reaction Temperature is now 37°C. Nb.BsrDI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10251908. 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 65°C in a total reaction volume of 50 μl. Reaction Conditions 1X rCutSmart™ Buffer Incubate at 37°C 1X rCutSmart™ Buffer 50 mM Potassium Acetate 20 mM Tris-acetate 10 mM Magnesium Acetate 100 µg/ml Recombinant Albumin (pH 7.9 @ 25°C) Activity in NEBuffers NEBuffer™ r1.1: 25% NEBuffer™ r2.1: 100% NEBuffer™ r3.1: 100% rCutSmart™ Buffer: 100% Diluent Compatibility Diluent A Storage Buffer 10 mM Tris-HCl 200 mM NaCl 1 mM DTT 0.1 mM EDTA 50% Glycerol 200 µg/ml Recombinant Albumin 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 FAQ 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. Q: Are certain restriction enzymes more active at higher incubation temperatures than what is recommended? A: NEB restriction enzymes are 100% active at the recommended incubation temperature in the recommended buffer provided with each enzyme. With some restriction enzymes isolated from thermophilic species, you may see increased activity at higher incubation temperatures. We do not recommend increasing the temperature as additional nuclease activities may appear at non-recommended temperatures.