NEBuffer Activity Chart for Restriction EnzymesNew 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 EnzymesThis chart has been updated to reflect the change of 54 restriction enzymes to NEBuffer 4 on Nov.1, 2024.Click for the Chart Legend, Icon DescriptionA | B | C | D | E | F | H | I | K | M | N | P | R | S | T | X | Z | EnzymeSupplied NEBuffer% Activity in NEBuffersHeatInactiva-tion °CIncuba.Temp.°CDiluent1234AatIINEBuffer 4050501006537AAcc65INEBuffer 3 + BSA1075100256537AAccINEBuffer 45050101008037AAciINEBuffer 32550100506537AAclINEBuffer 4 + BSA10100100No37BAcuINEBuffer 4 + SAM50100501006537BAfeINEBuffer 42550251006537AAflIINEBuffer 4 + BSA50100251006537AAflIIINEBuffer 3 + BSA2575100508037BAgeINEBuffer 11005010756537CAhdINEBuffer 4 + BSA257501006537AAleINEBuffer 41025101006537AAluINEBuffer 4100100751006537BAlwINEBuffer 450100101006537AAlwNINEBuffer 450100501006537AApaINEBuffer 4 + BSA255001006525AApaLINEBuffer 4 + BSA10010010100No37AApeKINEBuffer 3257510050No75BApoINEBuffer 3 + BSA1075100758050AAscINEBuffer 4010101006537AAseINEBuffer 3NR75100 ddNR6537BAsiSINEBuffer 4 + BSA501001001008037BAvaIN...