Isoschizomers Isoschizomers. The enzymes in boldface type are available from Promega. AatI StuI, Eco147I, Pme55I, SseBI AGGtCCT BsmI BsaMI GAATGCNt AatII — GACGTtC BsmAI1 Alw26I GTCTC(1/5) AccI FblI, XmiI GTt(A/C)(G/T)AC BsoBI AvaI, Ama87I, BcoI, Eco88I C(C/T)CG(G/A)G AccIII BspEI, MroI TtCCGGA Bsp19I NcoI CtCATGG Acc65I Asp718I GtGTACC Bsp68I NruI TCGtCGA KpnI* GGTACtC Bsp106I ClaI, BspDI ATtCGAT AccB1I BanI, BshNI, Eco64I GtG(C/T)(G/A)CC Bsp119I Csp45I, NspV, BstBI TTtCGAA t t AccB7I PflMI, Van91I CCAN4 NTGG Bsp120I — G GGCCC AclNI Spel AtCTAGT ApaI GGGCCtC AclWI AlwI GGATCNNNNt Bsp143I MboI, Sau3AI, NdeII tGATC AcyI BbiII, Hin1I, Hsp92I, G(A/G)tCG(T/C)C Bsp143II HaeII (A/G)GCGCt(T/C) BsaHI, Msp171I Bsp1286I BmyI, SduI G(G/A/T)GC(C/A/T)tC AcsI ApoI (G/A)tAATT(C/T) BspCI PvuI CGATtCG AfaI RsaI GTtAC BspDI ClaI ATtCGAT Csp6I GtTAC BspEI AccIII TtCCGGA AfeI Eco47III AGCtGCT BsrI1 BsrSI, BseNI ACTGGN(1/-1) AflII Bst98I CtTTAAG BsrSI1 BseNI, BsrI ACTGGN(1/-1) AgeI PinAI AtCCGGT BssHII BsePI, PauI GtCGCGC AhaIII DraI TTTtAAA Bst98I AflII, BfrI CtTTAAG AhdI EclHKI GACNNNtNNGTC BstBI Csp45I, NspV, Bsp119I TTtCGAA AluI — AGtCT BstEII BstPI, Eco91I, PspEI GtGTNACC AlwI AclWI GGATCNNNNt BstNI BstOI, MvaI CCt(A/T)GG Alw26I1 BsmAI GTCTC(1/5) EcoRII* tCC(A/T)GG Alw44I ApaLI GtTGCAC BstOI BstNI, MvaI CCt(A/T)GG AocI Bsu36I, CvnI CCtTNAGG EcoRII* tCC(A/T)GG ApaI — GGGCCtC BstXI – CCANNNNNtNTGG t t TECHNICAL REFERENCE Bsp120I* G GGCCC BstYI XhoII, MflI (A/G) GATC(T/C) ApaLI Alw44I, VneI GtTGCAC BstZI Eco52I, EagI, XmaIII, EclXI CtGGCCG ApoI AcsI (G/A)tAATT(C/T) Bsu15I ClaI ATtCGAT AseI VspI, AsnI ATtTAAT Bsu36I CvnI, AocI, Eco81I CCtTNAGG AsnI VspI, AseI ATtTAAT BsuRI HaeIII, PalI GGtCC AspI Tth111I GACNtNNGTC CfoI HhaI GCGtC AspEI AhdI, Eam1105I, EclHKI GACNNN/NNGTC Hin6I, HinP1I GtCGC Asp700I XmnI GAANNtNNTTC Cfr9I XmaI CtCCGGG Asp718I Acc65I GtGTACC SmaI* CCCtGGG KpnI* GGTACtC Cfr13I Sau96I GtGNCC AsuI Sau96I, Cfr13I GtGNCC Cfr42I SacII CCGCtGG AsuII Csp45I, BstBI TTtCGAA ClaI BanIII, Bsp106I, BspDI, Bsu15I ATtCGAT t t AsuHPI HphI GGTGAN8 CpoI CspI, RsrII CG G(A/T)CCG AvaI Ama87I, BcoI, BsoBI, Eco88I Ct(C/T)CG(G/A)G CspI CpoI, RsrII CGtG(A/T)CCG AvaII SinI, Eco47I, HgiEI GtG(A/T)CC Csp6I — GtTAC AxyI Bsu36I CCtTNAGG RsaI*, AfaI* GTtAC BalI MscI, MluNI TGGtCCA Csp45I BstBI, NspV, Bsp119I TTtCGAA BamHI — GtGATCC CvnI Bsu36I CCtTNAGG BanI AccBI, BshNI, Eco64I GtG(T/C)(A/G)CC DdeI BstDEI CtTNAG BanII Eco24I G(A/G)GC(T/C)tC DpnI2 — GmeAtTC BbeI — GGCGCtC DpnII* GAtTC NarI* GGtCGCC DpnII MboI, Sau3AI, NdeII, tGATC BbrPI Eco72I, PmlI CACtGTG DpnI* GAtTC BbsI1 Bsc91I, BpiI GAAGAC(2/6) DraI — TTTtAAA BbuI PaeI, SphI GCATGtC EagI Eco52I, BstZI, EclXI, XmaIII CtGGCCG BclI BsiQI, FbaI TtGATCA Eam1105I EclHKI, AhdI, AspEI GACNNNtNNGTC BcnI NciI CCt(C/G)GG Ecl136II EcoICRI GAGtCTC BfrI Bst98I CtTTAAG SacI* GAGCTtC BglI — GCCNNNNtNGGC EclHKI AhdI, Eam1105I, AspEI GACNNNtNNGTC BglII — AtGATCT EclXI BstZI, EagI, Eco52I, XmaIII CtGGCCG BmyI Bsp1286I G(G/A/T)GC(C/A/T)tC Eco24I BanII, FriOI G(AG)GC(TC)tC BpmI GsuI CTGGAG(16/14) Eco32I EcoRV GATtATC BsaHI Hsp92I G(A/G)tCG(T/C)C Eco47I AvaII, SinI GtG(A/T)CC BsaMI BsmI GAATGC(1/–1) Eco47III AfeI AGCtGCT BsaOI Bsh1285I, BsiEI CG(A/G)(T/C)tC Eco52I BstZI, XmaIII, EagI, EclXI CtGGCCG BseAI AccIII TtCCGGA Eco64I BanI, BshNI, Eco64I GtG(TC)(AG)CC BseNI BsrSI, BsrI ACTGGN (1/–1) Eco81I Bsu36I CCtTNAGG BsePI BssHII, PauI GtCGCGC Eco88I AvaI Ct(TC)CG(AG)G Bsh1285I BsaOI CG(A/G)(T/C)tCG Eco91I BstEII GtGTNACC BshNI BanI, AccB1I, Eco64I GtG(T/C)(A/G)CC Eco105I SnaBI TACtCTA Bsh1365I BsrBRI GATNNtNNATC BsiEI BsaOI CG(A/G)(T/C)tCG Isoschizomers Isoschizomers (continued). The enzymes in boldface type are available from Promega. Enzyme Isoschizomer(s) Recognition Sequence Enzyme Isoschizomer(s) Recognition Sequence Eco130I StyI CtC(A/T)(T/A)GG NspBII MspA1I C(A/C)GtC(G/T)G Eco147I StuI AGGtCCT PaeI BbuI, SphI GCATGtC EcoICRI Ecl136II GAGtCTC PaeR7I XhoI CtTCGAG SacI*, SstI* GAGCTtC PalI HaeIII, BsuRI GGtCC — GtAATTC t EcoRI PflMI AccB7I, Van91I CCAN4 NTGG EcoRII — tCC(A/T)GG PinAI AgeI AtCCGGT BstOI*, BstNI*, MvaI* CCt(A/T)GG PstI — CTGCAtG EcoRV Eco32I GATtATC PvuI BspCI CGATtCG EcoT14I StyI CtC(A/T)(A/T)GG PvuII — CAGtCTG EcoT22I NsiI ATGCAtT RsaI AfaI GTtAC EheI — GGCtGCC RsrII CspI, CpoI CGtG(A/T)CCG NarI* GGtCGCC SacI SstI GAGCTtC FokI2 — GGATG(9/13) Ecl136II*, EcoICRI* GAGtCTC HaeII Bsp143II (A/G)GCGCt(T/C) SacII SstII, KspI, Cfr42I CCGCtGG HaeIII BsuRI, PalI GGtCC SalI — GtTCGAC HapII HpaII, MspI CtCGG Sau3AI MboI, NdeII, DpnII tGATC HgiEI Eco47I, SinI, AvaII GtG(A/T)CC Sau96I Cfr13I GtGNCC HhaI CfoI GCGtC ScaI — AGTtACT HinP1I*, Hin6I* GtCGC SduI Bsp1286I G(G/A/T)GC(C/A/T)tC Hin1I AcyI, Hsp92I G(A/G)tCG(T/C)C SfiI — GGCCNNNNtNGGCC HincII HindII GT(T/C)t(A/G)AC SfuI Csp45I TTtCGAA HindII HincII GT(T/C)t(A/G)AC SgfI — GCGATtCGC HindIII — AtAGCTT SinI AvaII, Eco47I GtG(A/T)CC HinfI — GtANTC SmaI — CCCtGGG HinP1I — GtCGC XmaI*, Cfr9I* CtCCGGG HhaI*, CfoI* GCGtC SnaBI Eco105I TACtGTA TECHNICAL REFERENCEHpaI KspAI GTTtAAC SpeI AclNI AtCTAGT HpaII3 MspI, HapII CtCGG SphI BbuI, PaeI GCATGtC Hsp92I AcyI, BsaHI, Hin1I G(A/G)tCG(C/T)C SspI — AATtATT Hsp92II NlaIII CATGt SstI SacI GAGCTtC I-PpoI — CTCTCTTAAtGGTAGC EcoICRI* GAGtCTC KasI — GtGCGCC SstII SacII CCGCtGG NarI* GGtCGCC StuI AatI, Eco147I AGGtCCT KpnI — GGTACtC StyI EcoT14I CtC(A/T)(A/T)GG t Acc65I*, Asp718I* G GTACC TaqI TthHB8I TtCGA t KspI SacII CCGC GG Tru9I MseI TtTAA t MboI Sau3AI, NdeII, DpnII GATC Tth111I AspI GACNtNNGTC MboII1 — GAAGA(8/7) TthHB8I TaqI TtCGA t MflI XhoII (A/G) GATC(T/C) Van91I AccB7I, PflMI CCAN tNTGG t 4 MluI — A CGCGT VneI ApaLI, Alw44I GtTGCAC t MluNI BalI, MscI TGG CCA VspI AseI, AsnI ATtTAAT t MroI AccIII T CCGGA XbaI — TtCTAGA t MscI BalI, MluNI TGG CCA XhoI PaeR7I CtTCGAG t MseI Tru9I T TAA XhoII BstYI, MflI (A/G)tGATC(T/C) t MspI3 HpaII, HapII C CGG XmaI Cfr9I, XmaCI, CtCCGGG MspA1I NspBII C(A/C)GtC(G/T)G SmaI* CCCtGGG MstII Bsu36I CCtTNAGG XmaIII Eco52I, BstZI, EagI, EclXI CtGGCCG MvaI BstOI, BstNI CCt(A/T)GG XmaCI XmaI CtCCGGG EcoRII* tCC(A/T)GG SmaI* CCCtGGG NaeI — GCCtGGC XmnI Asp700I GAANNtNNTTC NgoMIV GtCCGGC NarI — GGtCGCC Key: EheI* GGCtGCC N = A, C, G or T KasI* GtGCGCC * = neoschizomer BbeI* GGCGCtC Notes: NciI BcnI CCt(C/G)GG 1. The locations of cleavage sites falling outside the recognition site are indicated in NcoI Bsp19I CtCATGG parentheses. For example, GTCTC(1/5) indicates cleavage at: 5′...GTCTCNt...3′ NdeI — CAtTATG 3′...CAGAGNNNNNs...5′ t NdeII MboI, Sau3AI, DpnII GATC 2. DpnI is unique among commercially available restriction enzymes in requiring methylation NgoMIV — GtCCGGC of a nucleotide (adenine) in its recognition sequence in order to cut. Therefore, DpnI t cannot be substituted for other enzymes recognizing the GATC sequence (e.g., MboI and NaeI* GCC GGC Sau3AI). t NheI — G CTAGC 3. Although HpaII and MspI recognize the same nucleotide sequence, HpaII is sensitive to NlaIII Hsp92II CATGt methylation of either cytosine in its recognition sequence, while MspI is sensitive only NotI — GCtGGCCGC to methylation of the external cytosine. These enzymes may not be interchanged for all applications. NruI Bsp68 I TCGtCGA NsiI EcoT22I, Mph1103I ATGCAtT Reference NspV Csp45I, BstBI, Bsp119I TTtCGAA Roberts, R.J. and Macelis, D. (1991) Nucl. Acids Res. 19 (supp), 2077. PART #GE633 PROMEGA CORPORATION 2800 WOODS HOLLOW ROAD MADISON, WI 53711-5399 USATELEPHONE 608-274-4330 www.promega.com ©2010 ALL RIGHTS RESERVED.