New England Biolabs, R0169L, Sau3AI

Sau3AI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10317346. Related Categories Restriction Endonucleases S Applications Restriction Enzyme Digestion Specification Unit Definition One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 μl. Reaction Conditions 1X NEBuffer™ r1.1 Incubate at 37°C 1X NEBuffer™ r1.1 10 mM Bis-Tris-Propane-HCl 10 mM MgCl2 100 µg/ml Recombinant Albumin (pH 7 @ 25°C) Activity in NEBuffers NEBuffer™ r1.1: 100% NEBuffer™ r2.1: 50% NEBuffer™ r3.1: 10% rCutSmart™ Buffer: 100% 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 65°C for 20 minutes Methylation Sensitivity dam methylation: Not Sensitive dcm methylation: Not Sensitive CpG Methylation: Blocked by Overlapping FAQ Q: What's the difference between DpnI, DpnII, MboI, and Sau3AI? A: They all recognize the same sequence but have different methylation sensitivities. DpnI will only cleave fully-adenomethylated dam sites and hemi-adenomethylated dam sites 60X more slowly. DpnII and MboI share methylation sensitivity and cleave dam sites which lack adenomethylation; each is blocked by complete dam methylation and probably by hemi-methylation as well. Sau3AI will cleave all dam sites regardless of adenomethylation but is completely blocked by methylated cytosines within the dam sequence. Q: Is this enzyme sensitive to dam, dcm or mammalian CpG methylation? A: Yes. This enzyme cannot cut recognition sites in mammalian gDNA that are blocked by overlapping 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.