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NEBuffer™ Activity/Performance Chart with Restriction Enzymes

NEB’s restriction enzyme buffer system makes your restriction digests easy and convenient. We are able to offer >210 restriction enzymes that cut in a single buffer, CutSmart® . This improves ease-of-use, especially when performing double digests.

In addition to indicating the performance of each enzyme in the 4 NEBuffers, the chart also indicates ligation and recutting, star activity, and whether or not more than 1-site is required for cleavage.
 

Legend

Not Sensitive   Impaired
Blocked Impaired by Overlapping
Blocked by Overlapping Impaired by Some Combinations of Overlapping
Blocked by Some Combinations of Overlapping

Please check out other technical reference information related to restriction enzymes:
Double Digestion | Heat Inactivation | Activity at 37°C | Diluent Buffers | Time-Saver Enzymes | High Fidelity (HF) Restriction Enzymes 

Chart Legend | Icon Descriptions

EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
AatII recombinant timesaver 5min cpgGACGT/CCutSmart® Buffer1050*5010080°C37°CNot SensitiveNot SensitiveBlockedλ DNA
AbaSI recombinantCutSmart® Buffer25505010065°C25°CNot SensitiveNot SensitiveNot SensitiveT4 wild-type phage DNA (fully ghmC-modified)e
Acc65I recombinant timesaver 5min dcm cpgG/GTACCNEBuffer 3.11075*1002565°C37°CNot SensitiveBlocked by Some Combinations of OverlappingBlocked by Some Combinations of OverlappingpBC4 DNA
AccI recombinant timesaver 5min cpgGT/MKACCutSmart® Buffer50501010080°C37°CNot SensitiveNot SensitiveBlocked by Overlappingλ DNA
AciI recombinant timesaver 5min cpgCCGC(-3/-1)CutSmart® Buffer102510010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA
AclI recombinant timesaver 5min cpgAA/CGTTCutSmart® Buffer101010100No37°CNot SensitiveNot SensitiveBlockedλ DNA
AcuI recombinant timesaver 5minCTGAAG(16/14)CutSmart® Buffer + SAM501005010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA1, b, d
AfeI recombinant cpgAGC/GCTCutSmart® Buffer251002510065°C37°CNot SensitiveNot SensitiveBlockedpXba DNA
AflII recombinant timesaver 5minC/TTAAGCutSmart® Buffer501001010065°C37°CNot SensitiveNot SensitiveNot SensitiveΦX174 RF I DNA
AflIII recombinantA/CRYGTNEBuffer 3.110501005080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
AgeI § recombinant cpgA/CCGGTNEBuffer 1.110075257565°C37°CNot SensitiveNot SensitiveBlockedλ DNA
AgeI-HF® recombinant timesaver 5min engineered cpgA/CCGGTCutSmart® Buffer100501010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA
AhdI recombinant timesaver 5min cpgGACNNN/NNGTCCutSmart® Buffer25251010065°C37°CNot SensitiveNot SensitiveImpaired by Some Combinations of Overlappingλ DNAa
AleI recombinant cpgCACNN/NNGTGCutSmart® Buffer10101010080°C37°CNot SensitiveNot SensitiveImpaired by Some Combinations of Overlappingλ DNA
AluI recombinant timesaver 5minAG/CTCutSmart® Buffer251005010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
AlwI recombinant damGGATC(4/5)CutSmart® Buffer505010100No37°CBlockedNot SensitiveNot Sensitiveλ DNA (dam-)1, b, d
AlwNI recombinant timesaver 5min dcmCAGNNN/CTGCutSmart® Buffer101005010080°C37°CNot SensitiveBlocked by OverlappingNot Sensitiveλ DNA
ApaI recombinant timesaver 5min dcm cpgGGGCC/CCutSmart® Buffer25251010065°C25°CNot SensitiveBlocked by OverlappingBlocked by OverlappingpXba DNA
ApaLI recombinant timesaver 5min cpgG/TGCACCutSmart® Buffer10010010100No37°CNot SensitiveNot SensitiveBlocked by Overlappingλ DNA (HindIII digest)
ApeKI recombinant timesaver 5min cpgG/CWGCNEBuffer 3.1255010010No75°CNot SensitiveNot SensitiveBlocked by Overlappingλ DNA
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
ApoI § recombinant timesaver 5minR/AATTYNEBuffer 3.110751007580°C50°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
ApoI-HF recombinant timesaver 5min engineeredR/AATTYCutSmart® Buffer101001010080°C37°CNot SensitiveNot SensitiveNot Sensitive
AscI recombinant timesaver 5min cpgGG/CGCGCCCutSmart® Buffer10101010080°C37°CNot SensitiveNot SensitiveBlockedλ DNA
AseI recombinant timesaver 5minAT/TAATNEBuffer 3.11050*1001065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3
AsiSI recombinant cpgGCGAT/CGCCutSmart® Buffer1001002510080°C37°CNot SensitiveNot SensitiveBlockedXhoI digested pXba2, b
AvaI recombinant timesaver 5min cpgC/YCGRGCutSmart® Buffer101002510080°C37°CNot SensitiveNot SensitiveBlockedλ DNA
AvaII recombinant timesaver 5min dcm cpgG/GWCCCutSmart® Buffer50751010080°C37°CNot SensitiveBlocked by OverlappingBlocked by Overlappingλ DNA
AvrII recombinant timesaver 5minC/CTAGGCutSmart® Buffer1005050100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA (HindIII digest)
BaeGI recombinant timesaver 5minGKGCM/CNEBuffer 3.175751002580°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BaeI recombinant timesaver 5min cpg(10/15)ACNNNNGTAYC(12/7)CutSmart® Buffer + SAM501005010065°C25°CNot SensitiveNot SensitiveBlocked by Some Combinations of OverlappingpXba DNAe
BamHI § recombinant timesaver 5minG/GATCCNEBuffer 3.175*100*100100*No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3
BamHI-HF® recombinant timesaver 5min engineeredG/GATCCCutSmart® Buffer1005010100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BanI recombinant dcm cpgG/GYRCCCutSmart® Buffer10251010065°C37°CNot SensitiveBlocked by Some Combinations of OverlappingBlocked by Some Combinations of Overlappingλ DNA1
BanII recombinantGRGCY/CCutSmart® Buffer1001005010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA2
BbsI § recombinant timesaver 5minGAAGAC(2/6)NEBuffer 2.1100100257565°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BbsI-HF® recombinant timesaver 5min engineeredGAAGAC(2/6)CutSmart® Buffer10101010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BbvCI recombinant cpgCCTCAGC(-5/-2)CutSmart® Buffer1010050100No37°CNot SensitiveNot SensitiveImpaired by Overlapping λ DNA1, a
BbvI recombinant timesaver 5minGCAGC(8/12)CutSmart® Buffer1001002510065°C37°CNot SensitiveNot SensitiveNot SensitivepBR322 (dcm+) DNA3
BccI recombinantCCATC(4/5)CutSmart® Buffer100501010065°C37°CNot SensitiveNot SensitiveNot SensitivepXba DNA3, b
BceAI recombinant cpgACGGC(12/14)NEBuffer 3.1100*100*100100*65°C37°CNot SensitiveNot SensitiveBlockedpBR322 (dcm+) DNA1
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
BcgI recombinant dam cpg(10/12)CGANNNNNNTGC(12/10)NEBuffer 3.1 + SAM1075*10050*65°C37°CImpaired by Overlapping Not SensitiveBlocked by Some Combinations of Overlappingλ DNAe
BciVI recombinant timesaver 5minGTATCC(6/5)CutSmart® Buffer100251010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
BclI recombinant timesaver 5min damT/GATCANEBuffer 3.15010010075No50°CBlockedNot SensitiveNot Sensitiveλ DNA (dam-)
BclI-HF recombinant timesaver 5min engineered damT/GATCACutSmart® Buffer1001001010065°C37°CBlockedNot SensitiveNot Sensitive3
BcoDI recombinant timesaver 5min cpgGTCTC(1/5)CutSmart® Buffer507575100No37°CNot SensitiveNot SensitiveImpaired by Some Combinations of Overlappingλ DNA
BfaI recombinantC/TAGCutSmart® Buffer10101010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA2, b
BfuAI recombinant timesaver 5min cpgACCTGC(4/8)NEBuffer 3.110251001065°C50°CNot SensitiveNot SensitiveImpaired by Overlapping λ DNA3
BfuCI timesaver 5min cpg/GATCCutSmart® Buffer100502510080°C37°CNot SensitiveNot SensitiveBlocked by Overlappingλ DNA
BglI recombinant timesaver 5min cpgGCCNNNN/NGGCNEBuffer 3.110251001065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
BglII recombinant timesaver 5minA/GATCTNEBuffer 3.1101010010No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BlpI recombinant timesaver 5minGC/TNAGCCutSmart® Buffer5010010100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAd
BmgBI recombinant timesaver 5min cpgCACGTC(-3/-3)NEBuffer 3.110101001065°C37°CNot SensitiveNot SensitiveBlockedλ DNA3, b, d
BmrI recombinantACTGGG(5/4)NEBuffer 2.17510075100*65°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA (HindIII digest)b
BmtI § recombinantGCTAG/CNEBuffer 3.110010010010065°C37°CNot SensitiveNot SensitiveNot SensitivepXba DNA2
BmtI-HF® recombinant timesaver 5min engineeredGCTAG/CCutSmart® Buffer501001010065°C37°CNot SensitiveNot SensitiveNot SensitivepXba DNA
BpmI recombinantCTGGAG(16/14)NEBuffer 3.17510010010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA2
Bpu10I recombinantCCTNAGC(-5/-2)NEBuffer 3.110251002580°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3, b, d
BpuEI recombinant timesaver 5minCTTGAG(16/14)CutSmart® Buffer + SAM50*10050*10065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAd
BsaAI recombinant timesaver 5min cpgYAC/GTRCutSmart® Buffer100100100100No37°CNot SensitiveNot SensitiveBlockedλ DNA
BsaBI recombinant dam cpgGATNN/NNATCCutSmart® Buffer501007510080°C60°CBlocked by OverlappingNot SensitiveBlocked by Some Combinations of Overlappingλ DNA (dam-)2
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
BsaHI recombinant timesaver 5min dcm cpgGR/CGYCCutSmart® Buffer5010010010080°C37°CNot SensitiveBlocked by Some Combinations of OverlappingBlockedλ DNA
BsaI § recombinant dcm cpgGGTCTC(1/5)CutSmart® Buffer75*7510010065°C37°CNot SensitiveImpaired by Some Combinations of OverlappingBlocked by Some Combinations of OverlappingpXba DNA3
BsaI-HF® recombinant timesaver 5min engineered dcm cpgGGTCTC(1/5)CutSmart® Buffer501002510065°C37°CNot SensitiveBlocked by OverlappingBlocked by Some Combinations of OverlappingpXba DNA
BsaJI recombinantC/CNNGGCutSmart® Buffer5010010010080°C60°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BsaWI recombinant timesaver 5minW/CCGGWCutSmart® Buffer101005010080°C60°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BsaXI timesaver 5min(9/12)ACNNNNNCTCC(10/7)CutSmart® Buffer50*100*10100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAe
BseRI recombinant timesaver 5minGAGGAG(10/8)CutSmart® Buffer100*1007510080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAd
BseYI recombinant cpgCCCAGC(-5/-1)NEBuffer 3.110501005080°C37°CNot SensitiveNot SensitiveBlocked by Overlappingλ DNAd
BsgI recombinant timesaver 5minGTGCAG(16/14)CutSmart® Buffer + SAM25502510065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAd
BsiEI recombinant timesaver 5min cpgCGRY/CGCutSmart® Buffer255010100No60°CNot SensitiveNot SensitiveBlockedλ DNA
BsiHKAI recombinantGWGCW/CCutSmart® Buffer25100100100No65°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BsiWI § recombinant timesaver 5min cpgC/GTACGNEBuffer 3.12550*1002565°C55°CNot SensitiveNot SensitiveBlockedΦX174 DNA
BsiWI-HF® recombinant timesaver 5min engineered cpgC/GTACGCutSmart® Buffer5010010100No37°CNot SensitiveNot SensitiveBlocked
BslI recombinant timesaver 5min dcm cpgCCNNNNN/NNGGCutSmart® Buffer5075100100No55°CNot SensitiveBlocked by Some Combinations of OverlappingBlocked by Some Combinations of Overlappingλ DNAb
BsmAI recombinant timesaver 5min cpgGTCTC(1/5)CutSmart® Buffer50100100100No55°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
BsmBI recombinant timesaver 5min cpgCGTCTC(1/5)NEBuffer 3.11050*1002580°C55°CNot SensitiveNot SensitiveBlockedλ DNA
BsmFI recombinant dcm cpgGGGAC(10/14)CutSmart® Buffer25505010080°C65°CNot SensitiveBlocked by OverlappingBlocked by OverlappingpBR322 (dcm+) DNA1
BsmI recombinant timesaver 5minGAATGC(1/-1)CutSmart® Buffer251001010080°C65°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BsoBI recombinant timesaver 5minC/YCGRGCutSmart® Buffer2510010010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
Bsp1286I recombinant timesaver 5minGDGCH/CCutSmart® Buffer25252510065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
BspCNI recombinant timesaver 5minCTCAG(9/7)CutSmart® Buffer + SAM100751010080°C25°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
BspDI recombinant dam cpgAT/CGATCutSmart® Buffer25755010080°C37°CBlocked by OverlappingNot SensitiveBlockedλ DNA
BspEI recombinant timesaver 5min dam cpgT/CCGGANEBuffer 3.110101001080°C37°CBlocked by OverlappingNot SensitiveImpairedλ DNA (dam-)
BspHI recombinant timesaver 5min damT/CATGACutSmart® Buffer10502510080°C37°CImpaired by Overlapping Not SensitiveNot Sensitiveλ DNA
BspMI recombinantACCTGC(4/8)NEBuffer 3.11050*1001065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BspQI recombinant timesaver 5minGCTCTTC(1/4)NEBuffer 3.110010010010080°C50°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3
BsrBI recombinant timesaver 5min cpgCCGCTC(-3/-3)CutSmart® Buffer5010010010080°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNAd
BsrDI recombinant timesaver 5minGCAATG(2/0)NEBuffer 2.110100752580°C65°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3, d
BsrFαI recombinant timesaver 5min engineered cpgR/CCGGYCutSmart® Buffer25250100No37°CNot SensitiveNot SensitiveBlocked
BsrFI recombinant cpgR/CCGGYCutSmart® Buffer10100*100*100No37°CNot SensitiveNot SensitiveBlockedpBR322 (dcm+) DNA1
BsrGI § recombinant timesaver 5minT/GTACANEBuffer 2.1251001002580°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BsrGI-HF® recombinant timesaver 5min engineeredT/GTACACutSmart® Buffer1010010010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BsrI timesaver 5minACTGG(1/-1)NEBuffer 3.110501001080°C65°CNot SensitiveNot SensitiveNot SensitiveΦX174 RF I DNAb
BssHII recombinant timesaver 5min cpgG/CGCGCCutSmart® Buffer10010010010065°C50°CNot SensitiveNot SensitiveBlockedλ DNA
BssKI recombinant timesaver 5min dcm cpg/CCNGGCutSmart® Buffer5010010010080°C60°CNot SensitiveBlocked by OverlappingBlocked by Overlappingλ DNAb
BssSαI recombinant timesaver 5min engineeredCACGAG(-5/-1)CutSmart® Buffer102510100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BssSI recombinantCACGAG(-5/-1)NEBuffer 3.1501001005080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BstAPI recombinant cpgGCANNNN/NTGCCutSmart® Buffer501002510080°C60°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNAb
BstBI recombinant timesaver 5min cpgTT/CGAACutSmart® Buffer7510010100No65°CNot SensitiveNot SensitiveBlockedλ DNA
BstEII  § recombinant timesaver 5minG/GTNACCNEBuffer 3.11075*10075*No60°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
BstEII-HF® recombinant timesaver 5min engineeredG/GTNACCCutSmart® Buffer101010100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BstNI recombinant timesaver 5minCC/WGGNEBuffer 3.11010010075No60°CNot SensitiveNot SensitiveNot Sensitiveλ DNAa
BstUI timesaver 5min cpgCG/CGCutSmart® Buffer5010025100No60°CNot SensitiveNot SensitiveBlockedλ DNAb
BstXI recombinant timesaver 5min dcmCCANNNNN/NTGGNEBuffer 3.110501002580°C37°CNot SensitiveBlocked by Some Combinations of OverlappingNot Sensitiveλ DNA3
BstYI recombinant timesaver 5minR/GATCYNEBuffer 2.12510075100No60°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BstZ17I § recombinant timesaver 5min cpgGTA/TACCutSmart® Buffer75100100100No37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA3, b
BstZ17I-HF® recombinant timesaver 5min engineered cpgGTATACCutSmart® Buffer10010010100No37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
Bsu36I recombinant timesaver 5minCC/TNAGGCutSmart® Buffer2510010010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA (HindIII digest)b
BtgI recombinant timesaver 5minC/CRYGGCutSmart® Buffer5010010010080°C37°CNot SensitiveNot SensitiveNot SensitivepBR322 (dcm+) DNA
BtgZI recombinant cpgGCGATG(10/14)CutSmart® Buffer10251010080°C60°CNot SensitiveNot SensitiveImpairedλ DNA3, b, d
BtsαI recombinant timesaver 5min engineeredGCAGTG(2/0)CutSmart® Buffer10010025100No55°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BtsCI recombinant timesaver 5minGGATG(2/0)CutSmart® Buffer101002510080°C50°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
BtsI recombinantGCAGTG(2/0)CutSmart® Buffer100505010080°C55°CNot SensitiveNot SensitiveNot Sensitiveλ DNA1, b
BtsIMutI recombinant engineeredCAGTG(2/0)CutSmart® Buffer100501010080°C55°CNot SensitiveNot SensitiveNot SensitivepUC19 DNAb
Cac8I timesaver 5min cpgGCN/NGCCutSmart® Buffer507510010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNAb
ClaI recombinant timesaver 5min dam cpgAT/CGATCutSmart® Buffer10505010065°C37°CBlocked by OverlappingNot SensitiveBlockedλ DNA (dam-)
CspCI recombinant timesaver 5min(11/13)CAANNNNNGTGG(12/10)CutSmart® Buffer + SAM101001010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAe
CviAII recombinant timesaver 5minC/ATGCutSmart® Buffer50501010065°C25°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
CviKI-1 recombinantRG/CYCutSmart® Buffer25100100100No37°CNot SensitiveNot SensitiveNot SensitivepBR322 (dcm+) DNA1, b
CviQI recombinant timesaver 5minG/TACNEBuffer 3.175100*10075*No25°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
DdeI recombinant timesaver 5minC/TNAGCutSmart® Buffer7510010010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
DpnI recombinant timesaver 5min cpgGA/TCCutSmart® Buffer1001007510080°C37°CNot SensitiveNot SensitiveBlocked by OverlappingpBR322 DNA (dam methylated)b
DpnII recombinant timesaver 5min dam/GATCNEBuffer DpnII2525100*2565°C37°CBlockedNot SensitiveNot Sensitiveλ DNA (dam-)
DraI recombinant timesaver 5minTTT/AAACutSmart® Buffer75755010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
DraIII § recombinant cpgCACNNN/GTGNEBuffer 3 + BSA25501002565°C37°CNot SensitiveNot SensitiveImpaired by Overlapping λ DNA
DraIII-HF® recombinant timesaver 5min engineered cpgCACNNN/GTGCutSmart® Buffer105010100No37°CNot SensitiveNot SensitiveImpaired by Overlapping λ DNAb
DrdI timesaver 5min cpgGACNNNN/NNGTCCutSmart® Buffer25501010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of OverlappingpUC19 DNA3
EaeI recombinant dcm cpgY/GGCCRCutSmart® Buffer10501010065°C37°CNot SensitiveBlocked by OverlappingBlocked by Overlappingλ DNAb
EagI § recombinant timesaver 5min cpgC/GGCCGNEBuffer 3.110251001065°C37°CNot SensitiveNot SensitiveBlockedpXba DNA
EagI-HF® recombinant timesaver 5min engineered cpgC/GGCCGCutSmart® Buffer2510010010065°C37°CNot SensitiveNot SensitiveBlockedpXba DNA
EarI recombinant timesaver 5min cpgCTCTTC(1/4)CutSmart® Buffer50101010065°C37°CNot SensitiveNot SensitiveImpaired by Overlapping λ DNAb, d
EciI recombinant cpgGGCGGA(11/9)CutSmart® Buffer100505010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA2
Eco53kI recombinant timesaver 5min cpgGAG/CTCCutSmart® Buffer1001001010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of OverlappingpXba DNA3, b
EcoNI recombinant timesaver 5minCCTNN/NNNAGGCutSmart® Buffer501007510065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
EcoO109I recombinant timesaver 5min dcmRG/GNCCYCutSmart® Buffer501005010065°C37°CNot SensitiveBlocked by OverlappingNot Sensitiveλ DNA (HindIII digest)3
EcoP15I recombinant timesaver 5minCAGCAG(25/27)NEBuffer 3.1 + ATP7510010010065°C37°CNot SensitiveNot SensitiveNot SensitivepUC19 DNAe
EcoRI § recombinant timesaver 5min cpgG/AATTCNEBuffer EcoRI25100*5050*65°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
EcoRI-HF® recombinant timesaver 5min engineered cpgG/AATTCCutSmart® Buffer101001010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
EcoRV § recombinant timesaver 5min cpgGAT/ATCNEBuffer 3.110501001080°C37°CNot SensitiveNot SensitiveImpaired by Some Combinations of Overlappingλ DNA
EcoRV-HF® recombinant timesaver 5min engineered cpgGAT/ATCCutSmart® Buffer2510010010065°C37°CNot SensitiveNot SensitiveImpaired by Some Combinations of Overlappingλ DNA
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
FatI recombinant/CATGNEBuffer 2.110100505080°C55°CNot SensitiveNot SensitiveNot SensitivepUC19 DNA
FauI recombinant cpgCCCGC(4/6)CutSmart® Buffer100501010065°C55°CNot SensitiveNot SensitiveBlockedλ DNA3, b, d
Fnu4HI recombinant timesaver 5min cpgGC/NGCCutSmart® Buffer101010100No37°CNot SensitiveNot SensitiveBlocked by Overlappingλ DNAa
FokI recombinant dcm cpgGGATG(9/13)CutSmart® Buffer1001007510065°C37°CNot SensitiveImpaired by Overlapping Impaired by Overlapping λ DNA3, b, d
FseI recombinant timesaver 5min dcm cpgGGCCGG/CCCutSmart® Buffer100751010065°C37°CNot SensitiveImpaired by Some Combinations of OverlappingBlockedAdenovirus-2 DNA
FspEI recombinantCC(12/16)CutSmart® Buffer10101010080°C37°CNot SensitiveNot SensitiveNot SensitivepBR322 (dcm+) DNAe
FspI recombinant timesaver 5min cpgTGC/GCACutSmart® Buffer1010010100No37°CNot SensitiveNot SensitiveBlockedλ DNAb
HaeII recombinant timesaver 5min cpgRGCGC/YCutSmart® Buffer251001010080°C37°CNot SensitiveNot SensitiveBlockedλ DNA
HaeIII recombinant timesaver 5minGG/CCCutSmart® Buffer501002510080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
HgaI recombinant cpgGACGC(5/10)NEBuffer 1.11001002510065°C37°CNot SensitiveNot SensitiveBlockedΦX174 DNA1
HhaI recombinant timesaver 5min cpgGCG/CCutSmart® Buffer2510010010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA
HincII recombinant timesaver 5min cpgGTY/RACNEBuffer 3.12510010010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
HindIII § recombinantA/AGCTTNEBuffer 2.125100505080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA2
HindIII-HF® recombinant timesaver 5min engineeredA/AGCTTCutSmart® Buffer101001010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
HinfI recombinant timesaver 5min cpgG/ANTCCutSmart® Buffer5010010010080°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
HinP1I recombinant timesaver 5min cpgG/CGCCutSmart® Buffer10010010010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA
HpaI recombinant cpgGTT/AACCutSmart® Buffer1075*25100No37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA1
HpaII recombinant timesaver 5min cpgC/CGGCutSmart® Buffer100501010080°C37°CNot SensitiveNot SensitiveBlockedλ DNA
HphI recombinant timesaver 5min dam dcmGGTGA(8/7)CutSmart® Buffer50501010065°C37°CBlockedBlockedNot Sensitiveλ DNAb, d
Hpy166II recombinant timesaver 5min cpgGTN/NACCutSmart® Buffer1001005010065°C37°CNot SensitiveNot SensitiveBlocked by OverlappingpBR322 DNA
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
Hpy188I recombinant damTCN/GACutSmart® Buffer251005010065°C37°CBlocked by OverlappingNot SensitiveNot SensitivepBR322 (dcm+) DNA1, b
Hpy188III recombinant dam cpgTC/NNGACutSmart® Buffer1001001010065°C37°CBlocked by OverlappingNot SensitiveBlocked by OverlappingpUC19 DNA3, b
Hpy99I recombinant cpgCGWCG/CutSmart® Buffer50101010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA
HpyAV recombinant timesaver 5min cpgCCTTC(6/5)CutSmart® Buffer1001002510065°C37°CNot SensitiveNot SensitiveImpaired by Overlapping λ DNA3, b, d
HpyCH4III recombinantACN/GTCutSmart® Buffer100251010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
HpyCH4IV recombinant timesaver 5min cpgA/CGTCutSmart® Buffer100502510065°C37°CNot SensitiveNot SensitiveBlockedpUC19 DNA
HpyCH4V recombinant timesaver 5minTG/CACutSmart® Buffer50502510065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
I-CeuI recombinantTAACTATAACGGTCCTAAGGTAGCGAA(-9/-13)CutSmart® Buffer10101010065°C37°CpBHS ScaI-linearized Control Plasmid
I-SceI recombinantTAGGGATAACAGGGTAAT(-9/-13)CutSmart® Buffer10502510065°C37°CpGPS2 NotI-linearized Control Plasmid
KasI recombinant cpgG/GCGCCCutSmart® Buffer501005010065°C37°CNot SensitiveNot SensitiveBlockedpBR322 DNA3
KpnI § recombinantGGTAC/CNEBuffer 1.1100751050*No37°CNot SensitiveNot SensitiveNot SensitivepXba DNA1
KpnI-HF® recombinant timesaver 5min engineeredGGTAC/CCutSmart® Buffer1002510100No37°CNot SensitiveNot SensitiveNot SensitivepXba DNA
LpnPI recombinantCCDG(10/14)CutSmart® Buffer10101010065°C37°CNot SensitiveNot SensitiveNot SensitivepBR322 (dcm+) DNAe
MboI recombinant timesaver 5min dam cpg/GATCCutSmart® Buffer7510010010065°C37°CBlockedNot SensitiveImpaired by Overlapping λ DNA (dam-)
MboII recombinant damGAAGA(8/7)CutSmart® Buffer100*1005010065°C37°CBlocked by OverlappingNot SensitiveNot Sensitiveλ DNA (dam-)b
MfeI § recombinantC/AATTGCutSmart® Buffer755010100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA2
MfeI-HF® recombinant timesaver 5min engineeredC/AATTGCutSmart® Buffer752510100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
MluCI recombinant timesaver 5min/AATTCutSmart® Buffer1001010100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
MluI § recombinant timesaver 5min cpgA/CGCGTNEBuffer 3.110501002580°C37°CNot SensitiveNot SensitiveBlockedλ DNA
MluI-HF® recombinant timesaver 5min engineered cpgA/CGCGTCutSmart® Buffer25100100100No37°CNot SensitiveNot SensitiveBlockedλ DNA
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
MlyI recombinant timesaver 5minGAGTC(5/5)CutSmart® Buffer50501010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb, d
MmeI recombinant timesaver 5min cpgTCCRAC(20/18)CutSmart® Buffer + SAM501005010065°C37°CNot SensitiveNot SensitiveBlocked by OverlappingΦX174 DNAb, c
MnlI recombinant timesaver 5minCCTC(7/6)CutSmart® Buffer751005010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
MscI recombinant dcmTGG/CCACutSmart® Buffer2510010010080°C37°CNot SensitiveBlocked by OverlappingNot Sensitiveλ DNA
MseI recombinant timesaver 5minT/TAACutSmart® Buffer751007510065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
MslI recombinant timesaver 5minCAYNN/NNRTGCutSmart® Buffer50501010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
MspA1I recombinant timesaver 5min cpgCMG/CKGCutSmart® Buffer10501010065°C37°CNot SensitiveNot SensitiveBlocked by Overlappingλ DNA
MspI recombinant timesaver 5minC/CGGCutSmart® Buffer7510050100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
MspJI recombinantCNNR(9/13)CutSmart® Buffer10101010065°C37°CNot SensitiveNot SensitiveNot SensitivepBR322 (dcm+) DNAe
MwoI recombinant timesaver 5min cpgGCNNNNN/NNGCCutSmart® Buffer10100100100No60°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
NaeI recombinant cpgGCC/GGCCutSmart® Buffer252510100No37°CNot SensitiveNot SensitiveBlockedpXba DNAb
NarI recombinant cpgGG/CGCCCutSmart® Buffer1001001010065°C37°CNot SensitiveNot SensitiveBlockedpXba DNA
Nb.BbvCI recombinantCCTCAGCCutSmart® Buffer2510010010080°C37°CNot SensitiveNot SensitiveNot Sensitivesupercoiled plasmid DNAe
Nb.BsmI recombinantGAATGCNEBuffer 3.110501001080°C65°CNot SensitiveNot SensitiveNot SensitivepBR322 DNAe
Nb.BsrDI recombinantGCAATGCutSmart® Buffer2510010010080°C65°CNot SensitiveNot SensitiveNot SensitivepUC19 DNAe
Nb.BssSI recombinantCACGAGNEBuffer 3.11010010025No37°CNot SensitiveNot SensitiveNot Sensitive
Nb.BtsI recombinantGCAGTGCutSmart® Buffer751007510080°C37°CNot SensitiveNot SensitiveNot SensitiveΦX174 DNAe
NciI recombinant timesaver 5min cpgCC/SGGCutSmart® Buffer1002510100No37°CNot SensitiveNot SensitiveImpaired by Overlapping λ DNAb
NcoI § recombinant timesaver 5minC/CATGGNEBuffer 3.110010010010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
NcoI-HF® recombinant timesaver 5min engineeredC/CATGGCutSmart® Buffer501001010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
NdeI recombinant timesaver 5minCA/TATGCutSmart® Buffer7510010010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
NgoMIV recombinant timesaver 5min cpgG/CCGGCCutSmart® Buffer1005010100No37°CNot SensitiveNot SensitiveBlockedAdenovirus-2 DNA1
NheI § recombinant timesaver 5min cpgG/CTAGCNEBuffer 2.11001001010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA (HindIII digest)
NheI-HF® recombinant timesaver 5min engineered cpgG/CTAGCCutSmart® Buffer100251010080°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA (HindIII digest)
NlaIII recombinant timesaver 5minCATG/CutSmart® Buffer10101010065°C37°CNot SensitiveNot SensitiveNot SensitiveΦX174 DNA
NlaIV recombinant dcm cpgGGN/NCCCutSmart® Buffer10101010065°C37°CNot SensitiveBlocked by OverlappingBlocked by OverlappingpBR322 DNA
NmeAIII recombinantGCCGAG(21/19)CutSmart® Buffer + SAM10101010065°C37°CNot SensitiveNot SensitiveNot SensitiveΦX174 RF I DNAc
NotI § recombinant timesaver 5min cpgGC/GGCCGCNEBuffer 3.110501002565°C37°CNot SensitiveNot SensitiveBlockedpBC4 DNA
NotI-HF® recombinant timesaver 5min engineered cpgGC/GGCCGCCutSmart® Buffer251002510065°C37°CNot SensitiveNot SensitiveBlockedpBC4 DNA
NruI § recombinant timesaver 5min dam cpgTCG/CGANEBuffer 3.1101010010No37°CBlocked by OverlappingNot SensitiveBlockedλ DNAb
NruI-HF® recombinant timesaver 5min engineered dam cpgTCG/CGACutSmart® Buffer02550100No37°CBlocked by OverlappingNot SensitiveBlockedλ DNA
NsiI § recombinant timesaver 5minATGCA/TNEBuffer 3.110751002565°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
NsiI-HF® recombinant timesaver 5min engineeredATGCA/TCutSmart® Buffer10201010080°C37°CNot SensitiveNot SensitiveNot Sensitive
NspI recombinant timesaver 5minRCATG/YCutSmart® Buffer1001001010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
Nt.AlwI recombinant damGGATC(4/-5)CutSmart® Buffer1010010010080°C37°CBlockedNot SensitiveNot SensitivepUC101 DNA (dam-/dcm-)e
Nt.BbvCI recombinant cpgCCTCAGC(-5/-7)CutSmart® Buffer501001010080°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingsupercoiled plasmid DNAe
Nt.BsmAI recombinant cpgGTCTC(1/-5)CutSmart® Buffer100501010065°C37°CNot SensitiveNot SensitiveBlockedsupercoiled plasmid DNAe
Nt.BspQI recombinantGCTCTTC(1/-7)NEBuffer 3.110251001080°C50°CNot SensitiveNot SensitiveNot SensitivepUC19 DNAe
Nt.BstNBI recombinantGAGTC(4/-5)NEBuffer 3.10101001080°C55°CNot SensitiveNot SensitiveNot SensitiveT7 DNA
Nt.CviPII recombinant cpg(0/-1)CCDCutSmart® Buffer101002510065°C37°CNot SensitiveNot SensitiveBlockedpUC19 DNAe
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
PacI recombinant timesaver 5minTTAAT/TAACutSmart® Buffer100751010065°C37°CNot SensitiveNot SensitiveNot SensitivepNEB193 DNA
PaeR7I recombinant timesaver 5min cpgC/TCGAGCutSmart® Buffer2510010100No37°CNot SensitiveNot SensitiveBlockedλ DNA (HindIII digest)
PciI recombinantA/CATGTNEBuffer 3.1507510050*80°C37°CNot SensitiveNot SensitiveNot SensitivepXba DNA
PflFI recombinant timesaver 5minGACN/NNGTCCutSmart® Buffer251002510065°C37°CNot SensitiveNot SensitiveNot SensitivepBC4 DNAb
PflMI recombinant timesaver 5min dcmCCANNNN/NTGGNEBuffer 3.101001005065°C37°CNot SensitiveBlocked by OverlappingNot Sensitiveλ DNA3, b, d
PhoI recombinant dcm cpgGG/CCCutSmart® Buffer5050100100No75°CNot SensitiveImpaired by Some Combinations of OverlappingImpaired by Some Combinations of Overlappingλ DNA
PI-PspI recombinantTGGCAAACAGCTATTATGGGTATTATGGGT(-13/-17)NEBuffer PI-PspI + BSA10101010No65°CpAKR7 XmnI-linearized Control Plasmid
PI-SceI recombinantATCTATGTCGGGTGCGGAGAAAGAGGTAAT(-15/-19)NEBuffer PI-SceI + BSA1010101065°C37°CpBSvdeX XmnI-linearized Control Plasmid
PleI recombinant cpgGAGTC(4/5)CutSmart® Buffer25502510065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNAb, d
PluTI recombinant cpgGGCGC/CCutSmart® Buffer100251010065°C37°CNot SensitiveNot SensitiveBlockedpXba DNAb
PmeI recombinant timesaver 5min cpgGTTT/AAACCutSmart® Buffer10501010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
PmlI recombinant timesaver 5min cpgCAC/GTGCutSmart® Buffer100501010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA (HindIII digest)
PpuMI recombinant timesaver 5min dcmRG/GWCCYCutSmart® Buffer101010100No37°CNot SensitiveBlocked by OverlappingNot Sensitiveλ DNA (HindIII digest)
PshAI recombinant timesaver 5min cpgGACNN/NNGTCCutSmart® Buffer25501010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
PsiI recombinantTTA/TAACutSmart® Buffer101001010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3
PspGI recombinant dcm/CCWGGCutSmart® Buffer2510050100No75°CNot SensitiveBlockedNot SensitiveT7 DNA3
PspOMI recombinant dcm cpgG/GGCCCCutSmart® Buffer10101010065°C37°CNot SensitiveImpaired by Some Combinations of OverlappingBlocked by OverlappingpXba DNA
PspXI recombinant cpgVC/TCGAGBCutSmart® Buffer1010025100No37°CNot SensitiveNot SensitiveImpairedλ DNA (HindIII digest)
PstI § recombinant timesaver 5minCTGCA/GNEBuffer 3.1757510050*80°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
PstI-HF® recombinant timesaver 5min engineeredCTGCA/GCutSmart® Buffer107550100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
PvuI § recombinant timesaver 5min cpgCGAT/CGNEBuffer 3.1102510010No37°CNot SensitiveNot SensitiveBlockedpXba DNA
PvuI-HF® recombinant timesaver 5min engineered cpgCGAT/CGCutSmart® Buffer25100100100No37°CNot SensitiveNot SensitiveBlockedpXba DNA
PvuII § recombinant timesaver 5minCAG/CTGNEBuffer 3.150100100100*No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
PvuII-HF® recombinant timesaver 5min engineeredCAG/CTGCutSmart® Buffer101010100No37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
RsaI recombinant timesaver 5min cpgGT/ACCutSmart® Buffer255010100No37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
RsrII recombinant cpgCG/GWCCGCutSmart® Buffer25751010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA
SacI § recombinant timesaver 5minGAGCT/CNEBuffer 1.1100501010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA (HindIII digest)
SacI-HF® recombinant timesaver 5min engineered cpgGAGCT/CCutSmart® Buffer10501010065°C37°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA (HindIII digest)
SacII recombinant timesaver 5min cpgCCGC/GGCutSmart® Buffer101001010065°C37°CNot SensitiveNot SensitiveBlockedpXba DNA
SalI § recombinant timesaver 5min cpgG/TCGACNEBuffer 3.110101001065°C37°CNot SensitiveNot SensitiveBlockedλ DNA (HindIII digest)
SalI-HF® recombinant timesaver 5min engineered cpgG/TCGACCutSmart® Buffer1010010010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA (HindIII digest)
SapI recombinant timesaver 5minGCTCTTC(1/4)CutSmart® Buffer75501010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
Sau3AI recombinant cpg/GATCNEBuffer 1.1100501010065°C37°CNot SensitiveNot SensitiveBlocked by Overlappingλ DNAb
Sau96I recombinant dcm cpgG/GNCCCutSmart® Buffer5010010010065°C37°CNot SensitiveBlocked by OverlappingBlocked by Overlappingλ DNA
SbfI § recombinant timesaver 5minCCTGCA/GGCutSmart® Buffer50251010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3
SbfI-HF® recombinant timesaver 5min engineeredCCTGCA/GGCutSmart® Buffer50251010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
ScaI § recombinantAGT/ACTNEBuffer 3N/RN/R100N/R80°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
ScaI-HF® recombinant timesaver 5min engineeredAGT/ACTCutSmart® Buffer1001001010080°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
ScrFI recombinant dcm cpgCC/NGGCutSmart® Buffer10010010010065°C37°CNot SensitiveBlocked by OverlappingBlocked by Overlappingλ DNA2, a
SexAI recombinant dcmA/CCWGGTCutSmart® Buffer100755010065°C37°CNot SensitiveBlockedNot SensitivepBC4 DNA (dcm-)3, b, d
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
SfaNI recombinant cpgGCATC(5/9)NEBuffer 3.110751002565°C37°CNot SensitiveNot SensitiveImpaired by Some Combinations of OverlappingΦX174 RF I DNA3, b
SfcI recombinantC/TRYAGCutSmart® Buffer75502510065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA3
SfiI recombinant timesaver 5min dcm cpgGGCCNNNN/NGGCCCutSmart® Buffer2510050100No50°CNot SensitiveImpaired by Overlapping Blocked by Some Combinations of OverlappingAdenovirus-2 DNA
SfoI recombinant timesaver 5min cpg dcmGGC/GCCCutSmart® Buffer50100100100No37°CNot SensitiveBlocked by Some Combinations of OverlappingBlockedλ DNA (HindIII digest)
SgrAI recombinant cpgCR/CCGGYGCutSmart® Buffer1001001010065°C37°CNot SensitiveNot SensitiveBlockedλ DNA1
SmaI recombinant timesaver 5min cpgCCC/GGGCutSmart® Buffer10101010065°C25°CNot SensitiveNot SensitiveBlockedλ DNA (HindIII digest)b
SmlI recombinantC/TYRAGCutSmart® Buffer257525100No55°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
SnaBI recombinant cpgTAC/GTACutSmart® Buffer50*501010080°C37°CNot SensitiveNot SensitiveBlockedT7 DNA1
SpeI § recombinant timesaver 5minA/CTAGTCutSmart® Buffer751002510080°C37°CNot SensitiveNot SensitiveNot SensitiveAdenovirus-2 DNA
SpeI-HF® recombinant timesaver 5min engineeredA/CTAGTCutSmart® Buffer25501010080°C37°CNot SensitiveNot SensitiveNot SensitivepXba DNA
SphI § recombinantGCATG/CNEBuffer 2.11001005010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA2
SphI-HF® recombinant timesaver 5min engineeredGCATG/CCutSmart® Buffer50251010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
SrfI recombinant timesaver 5min engineered cpgGCCC/GGGCCutSmart® Buffer1050010065°C37°CNot SensitiveNot SensitiveBlocked
SspI § recombinant timesaver 5minAAT/ATTNEBuffer SspI50100505065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
SspI-HF® recombinant timesaver 5min engineeredAAT/ATTCutSmart® Buffer251001010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
StuI recombinant timesaver 5min dcmAGG/CCTCutSmart® Buffer5010050100No37°CNot SensitiveBlocked by OverlappingNot Sensitiveλ DNA
StyD4I recombinant timesaver 5min dcm cpg/CCNGGCutSmart® Buffer1010010010065°C37°CNot SensitiveBlocked by OverlappingImpaired by Overlapping λ DNA
StyI § recombinant timesaver 5minC/CWWGGNEBuffer 3.110251001065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
StyI-HF® recombinant timesaver 5min engineeredC/CWWGGCutSmart® Buffer251002510065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
SwaI recombinant timesaver 5minATTT/AAATNEBuffer 3.110101001065°C25°CNot SensitiveNot SensitiveNot SensitiveM13mp19 RF I DNAb, d
EnzymeSequenceSupplied NEBuffer% Activity in NEBufferHeat Inac.Incu. Temp.DiluentDamDcmCpGUnit SubstrateNote
1.12.13.1CutSmart
TaqαI recombinant timesaver 5min damT/CGACutSmart® Buffer507510010080°C65°CBlocked by OverlappingNot SensitiveNot Sensitiveλ DNA
TfiI recombinant timesaver 5min cpgG/AWTCCutSmart® Buffer50100100100No65°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA
TliI recombinant cpgC/TCGAGNEBuffer 3 + BSA105010050No75°CNot SensitiveNot SensitiveImpairedλ DNA (HindIII digest)
TseI timesaver 5min cpgG/CWGCCutSmart® Buffer75100100100No65°CNot SensitiveNot SensitiveBlocked by Some Combinations of Overlappingλ DNA3
Tsp45I/GTSACCutSmart® Buffer1005010100No65°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
Tsp509I/AATTNEBuffer 1N/RN/RN/RN/RNo65°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
TspMI timesaver 5min cpgC/CCGGGCutSmart® Buffer50*75*50*100No75°CNot SensitiveNot SensitiveBlockedpUCAdeno plasmid DNAd
TspRI recombinant timesaver 5minNNCASTGNN/CutSmart® Buffer255025100No65°CNot SensitiveNot SensitiveNot Sensitiveλ DNA
Tth111I recombinant timesaver 5minGACN/NNGTCCutSmart® Buffer2510025100No65°CNot SensitiveNot SensitiveNot SensitivepBC4 DNAb
XbaI recombinant timesaver 5min damT/CTAGACutSmart® Buffer101007510065°C37°CBlocked by OverlappingNot SensitiveNot Sensitiveλ DNA (dam-/Hind III digest)
XcmI recombinantCCANNNNN/NNNNTGGNEBuffer 2.1101002510065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNA2
XhoI recombinant timesaver 5min cpgC/TCGAGCutSmart® Buffer7510010010065°C37°CNot SensitiveNot SensitiveImpairedλ DNA (HindIII digest)b
XmaI recombinant timesaver 5min cpgC/CCGGGCutSmart® Buffer25501010065°C37°CNot SensitiveNot SensitiveImpairedAdenovirus-2 DNA3
XmnI recombinant timesaver 5minGAANN/NNTTCCutSmart® Buffer50751010065°C37°CNot SensitiveNot SensitiveNot Sensitiveλ DNAb
ZraI recombinant cpgGAC/GTCCutSmart® Buffer100251010080°C37°CNot SensitiveNot SensitiveBlockedλ DNA
Ligation and Recutting Notes
a. Ligation is less than 10%.
b. Ligation is 25% -75%.
c. Recutting after ligation is less than 5%.
d. Recutting after ligation is 50% -75%.
e. Ligation and recutting after ligation is not applicable since the enzyme is either a nicking enzyme, is affected by methylation, or the recognition sequence contains variable sequences.

Star Activity Notes
1. Star activity may result from extended digestion, high enzyme concentration or a glycerol concentration of > 5%.
2. Star activity may result from extended digestion.
3. Star activity may result from a glycerol concentration of > 5%.
*. May exhibit star activity in this buffer.

§ A HF version of this enzyme is available.
The values listed in this table are approximate. They were obtained using each enzyme's specific unit assay substrate DNA.