New England Biolabs, R0607L, Nt.BstNBI

Nt.BstNBI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10159526. Related Categories Nicking Endonucleases,, Restriction Endonucleases: N-O Applications Strand Displacement Amplification & Nicking Enzyme Specification Unit Definition One unit is defined as the amount of enzyme required to digest 1 µg T7 DNA in 1 hour at 55°C in a total reaction volume of 50 μl. The phage T7 DNA is a particularly convenient substrate for assaying the activity because of the proximity of GAGTC sequences. Nt.BstNBI cleavage of phage T7 DNA yields 5 bands; 13 kb, 9 kb, 7 kb, 6 kb and 4 kb. Reaction Conditions 1X NEBuffer™ r3.1 Incubate at 55°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: 0% NEBuffer™ r2.1: 10% NEBuffer™ r3.1: 100% rCutSmart™ Buffer: 10% Diluent Compatibility Diluent A Storage Buffer 10 mM Tris-HCl 50 mM KCl 1 mM DTT 0.1 mM EDTA 200 µg/ml Recombinant Albumin 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: What is the activity of nicking enzymes at different temperatures? A: The activity of NEB nicking enzymes at different temperatures can be found on the table "Effect of Various Temperatures on Nicking Endonucleases”. 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.