New England Biolabs, R0650L, FatI

FatI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10201943. Related Categories Restriction Endonucleases C G Applications Restriction Enzyme Digestion Specification Unit Definition One unit is defined as the amount of enzyme required to digest 1 µg of pUC19 DNA in 1 hour at 55°C in a total reaction volume of 50 μl. Reaction Conditions 1X NEBuffer™ r2.1 Incubate at 55°C 1X NEBuffer™ r2.1 50 mM NaCl 10 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: 100% NEBuffer™ r3.1: 50% rCutSmart™ Buffer: 50% Diluent Compatibility Diluent A Storage Buffer 10 mM Tris-HCl 50 mM NaCl 0.1 mM EDTA 1 mM DTT 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 Activity at Temperature @37°C: 100% FAQ Q: How many base pairs should be added at the end of a PCR primer after the FatI recognition site to guarantee that FatI will cut properly? A: The addition of 6 base pairs is recommended. Q: Does FatI have any neoschizomers? A: Yes. CviAII and NlaIII are neoschizomer of FatI. For the most current information about isoschizomers/neoschizomers please refer to REBASE. 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.