NEBuffer Activity Chart for Restriction Enzymes New England Biolabs provides a color-coded 10X NEBuffer with every restriction endonuclease to ensure 100% activity. To help select the best conditions for double digests, this chart shows the optimal (supplied) NEBuffer and approximate activity in the four standard NEBuffers for each enzyme. If BSA is supplied with an enzyme, it is included in all NEBuffer activity reactions. In addition, the table also shows recommended reaction temperature, heat inactivation temperature and recommended diluent buffer. Please click here for additional information about: Unique NEBuffer Activity | Double Digestion | Heat Inactivation | Activity at 37°C | Diluent Buffers | Time-Saver Enzymes | High Fidelity (HF) Restriction Enzymes This chart has been updated to reflect the change of 54 restriction enzymes to NEBuffer 4 on Nov.1, 2008. Click for the Chart Legend, Icon Description A | B | C | D | E | F | H | I | K | M | N | P | R | S | T | X | Z | Enzyme Supplied NEBuffer % Activity in NEBuffers Heat Inactiva- tion °C Incuba. Temp. °C Diluent 1 2 3 4 AatII NEBuffer 4 0 50 50 100 65 37 A Acc65I NEBuffer 3 + BSA 10 75 100 25 65 37 A AccI NEBuffer 4 50 50 10 100 80 37 A AciI NEBuffer 3 25 50 100 50 65 37 A AclI NEBuffer 4 + BSA 10 10 0 100 No 37 B AcuI NEBuffer 4 + SAM 50 100 50 100 65 37 B AfeI NEBuffer 4 25 50 25 100 65 37 A AflII NEBuffer 4 + BSA 50 100 25 100 65 37 A AflIII NEBuffer 3 + BSA 25 75 100 50 80 37 B AgeI NEBuffer 1 100 50 10 75 65 37 C AhdI NEBuffer 4 + BSA 25 75 0 100 65 37 A AleI NEBuffer 4 10 25 10 100 65 37 A AluI NEBuffer 4 100 100 75 100 65 37 B AlwI NEBuffer 4 50 100 10 100 65...
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