New England Biolabs, R0540L, BspEI

BspEI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10192518. Related Categories Restriction Endonucleases B,, Time-Saver Qualified Restriction Enzymes Applications Restriction Enzyme Digestion Specification Unit Definition One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA (dam-) in 1 hour at 37°C in a total reaction volume of 50 μl. Reaction Conditions 1X NEBuffer™ r3.1 Incubate at 37°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: 10% 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 Recombinant Albumin 50% Glycerol pH 7.4 @ 25°C Heat Inactivation 80°C for 20 minutes Methylation Sensitivity dam methylation: Blocked by Overlapping dcm methylation: Not Sensitive CpG Methylation: Impaired FAQ Q: Is BspEI activity sensitive to dam, dcm or mammalian CpG methylation? A: Yes. BspEI cannot cut recognition sites that contain overlapping dam methylation. In mammalian gDNA BspEI cannot cut sites that contain CpG methylation. For up-to-date information about methylation sensitivities, please visit Dam-Dcm and CpG Methylation or REBASE. Q: How many base pairs should be added at the end of a PCR primer next to the BspEI recognition site to guarantee that BspEI will cut properly? A: The addition of 6 base pairs is recommended. Q: Does BspEI have any neoschizomers? A: No. For the most current information about isoschizomers/neoschizomers please refer to REBASE. Q: Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer? A: NEB's Time-Saver™ enzymes have the benefit of working fast (5-15 minutes), but are also designed and qualified to withstand overnight digestions without degradation of DNA. 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.