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NEBufferActivityChartforRestrictionEnzymesNewEnglandBiolabsprovidesacolor-coded10XNEBufferwitheveryrestrictionendonucleasetoensure100%activity.Tohelpselectthebestconditionsfordoubledigests,thischartshowstheoptimal(supplied)NEBufferandapproximateactivityinthefourstandardNEBuffersforeachenzyme.IfBSAissuppliedwithanenzyme,itisincludedinallNEBufferactivityreactions.Inaddition,thetablealsoshowsrecommendedreactiontemperature,heatinactivationtemperatureandrecommendeddiluentbuffer.Pleaseclickhereforadditionalinformationabout:

UniqueNEBufferActivity

|DoubleDigestion|HeatInactivation|Activityat37°C|DiluentBuffers|Time-SaverEnzymes|HighFidelity(HF)RestrictionEnzymesThischarthasbeenupdatedtoreflectthechangeof54restrictionenzymestoNEBuffer4onNov.1,2023.ClickfortheChartLegend,IconDescriptionA|B|C|D|E|F|H|I|K|M|N|P|R|S|T|X|Z|EnzymeSuppliedNEBuffer%ActivityinNEBuffersHeat

Inactiva-

tion°CIncuba.

Temp.

°CDiluent1234AatIINEBuffer4050501006537AAcc65INEBuffer3+BSA1075100256537AAccINEBuffer45050101008037AAciINEBuffer32550100506537AAclINEBuffer4+BSA10100100No37BAcuINEBuffer4+SAM50100501006537BAfeINEBuffer42550251006537AAflIINEBuffer4+BSA50100251006537AAflIIINEBuffer3+BSA2575100508037BAgeINEBuffer11005010756537CAhdINEBuffer4+BSA257501006537AAleINEBuffer41025101006537AAluINEBuffer4100100751006537BAlwINEBuffer450100101006537AAlwNINEBuffer450100501006537AApaINEBuffer4+BSA255001006525AApaLINEBuffer4+BSA10010010100No37AApeKINEBuffer3257510050No75BApoINEBuffer3+BSA1075100758050AAscINEBuffer4010101006537AAseINEBuffer3NR75100ddNR6537BAsiSINEBuffer4+BSA501001001008037BAvaINEBuffer41075101008037AAvaIINEBuffer45075101006537AAvrIINEBuffer4100100501008037BEnzyme

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentBaeGINEBuffer11007510506537ABaeINEBuffer4+BSA+SAM50100501006525ABamHINEBuffer3+BSA75100100dd100No37ABamHI-HF™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

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentCac8INEBuffer450751001006537BClaINEBuffer4+BSA1050501006537ACspCINEBuffer4+SAM10100101006537ACviAIINEBuffer47525101006525BCviKI-1NEBuffer41050251008037ACviQINEBuffer3+BSA7510010075No25BEnzyme

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentDdeINEBuffer375100100756537ADpnINEBuffer4100100751008037BDpnIINEBufferDpnIINRNR100ddNR6537BDraINEBuffer47575501006537ADraIIINEBuffer3+BSA10075100256537BDrdINEBuffer4+BSA2550101006537AEnzyme

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentEaeINEBuffer4100100501008037AEagINEBuffer31025100106537CEagI-HF™NEBuffer42550101006537CEarINEBuffer4100100501006537AEciINEBuffer2+BSA50100100506537AEcoNINEBuffer4100100751006537AEcoO109INEBuffer4+BSA100100751006537AEcoP15INEBuffer3+BSA+ATP751001001006537AEcoRINEBufferEcoRIdd1001001001006537CEcoRI-HF™NEBuffer41010001006537CEcoRVNEBuffer3+BSA5075100508037AEcoRV-HF™NEBuffer4251001001006537BEnzyme

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentFatINEBuffer21010050506555AFauINEBuffer410050101006555AFnu4HINEBuffer41025251006537AFokINEBuffer4100100751006537AFseINEBuffer4+BSA1007501006537BFspINEBuffer4107510100dd6537CEnzyme

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentHaeIINEBuffer4+BSA75100501008037AHaeIIINEBuffer450100251008037AHgaINEBuffer110075501006537AHhaINEBuffer4+BSA751001001006537AHincIINEBuffer3+BSA751001001006537BHindIIINEBuffer25010010506537BHinfINEBuffer475100751008037AHinP1INEBuffer41001001001006537AHpaINEBuffer4255010100No37AHpaIINEBuffer11005010506537AHphINEBuffer4NR7501006537BHpy166IINEBuffer4100100501006537CHpy188INEBuffer45025101006537AHpy188IIINEBuffer4+BSA100100101006537AHpy99INEBuffer4+BSA101001006537AHpyAVNEBuffer4+BSA100100251006537BHpyCH4IIINEBuffer410050251008037AHpyCH4IVNEBuffer11002510256537AHpyCH4VNEBuffer45050251006537BEnzyme

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentI-CeuINEBuffer4+BSA101001006537BI-SceINEBufferI-SceI+BSA105050506537BEnzyme

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentKasINEBuffer4+BSA2510001006537BKpnINEBuffer1+BSA10075050No37AEnzyme

topSuppliedNEBuffer%inB1%inB2%inB3%inB4Heat

Inactiva.Incuba.

Temp.DiluentMboINEBuffer4751001001006537AMboIINEBuffer4100100501006537CMfeINEBuffer47550101006537AMfeI-HF™NEBuffer47550101006537AMluINEBuffer32575100506537AMlyINEBuffer4+BSA5050251006537AMmeINEBuffer4+SAMNRNRNR100dd8037BMnlINEBuffer4+BSA75100501006537BMscINEBuffer47575751006537BMseINEBuffer4+BSA75100751006537AMslINEBuffer4100100251006537AMspA1INEBuffer4+BSA50100501006537BMspINEBuffer

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