TECHNICAL REFERENCE BsaHI, AlwI AcyI AclWI AclNI AccB7I AccB1I Acc65I AccIII AccI AatII AatI Isoschizomers. sE BsaOI BsiEI BsrBRI Bsh1365I BshNI BsaOI Bsh1285I BsePI BseNI Bsh1285I, BseAI BsaOI GsuI BsaMI BsaHI BpmI BmyI BglII BglI BfrI BcnI BclI BbuI Eco72I, BbsI — BbrPI BbeI BanII BanI BamHI BalI AxyI AvaII HphI AvaI AsuHPI AsuII AsuI Asp718I AhdI, Asp700I AspEI AspI AcsI AsnI Bsp120I* AseI ApoI ApaLI ApaI AocI Alw44I Alw26I AlwI AluI AhdI AhaIII Csp6I AgeI AflII AfeI ApoI AfaI AcsI 1 BsiQI, MscI, — AclWI BbiII,

PinAI Sau96I, AccBI, — Bsc91I, — PaeI, — —

Ama87I, FblI,

Eco24I 1 BsmI BspEI,

Asp718I PflMI, — BsmAI ApaLI

BanI BssHII BsrSI AccIII Hsp92I Bsp1286I Bst98I NciI NarI Bsu36I SinI Csp45I KpnI Acc65I XmnI Tth111I VspI VspI Alw44I Bsu36I EclHKI DraI Bst98I Eco47III RsaI Spel BanI KpnI StuI ,

, * , ,

XmiI * , * , Hin1I, SphI Eco47I,

Eam1105I, Eco147I, , MluNI AseI AsnI

Van91I BshNI, AccB1I, FbaI MroI BshNI, , The enzymes in boldface type are available from Promega. from available are type boldface in enzymes The Msp171I

BsrI ,

, ,

PmlI BpiI

Cfr13I PauI

BstBI VneI CvnI BcoI,

BsiEI Hsp92I Eco64I HgiEI Eco64I BsoBI, Eco64I Pme55I, EclHKI , c8I C Eco88I sB AGG SseBI GACNNN/NNGTC GAAGAC(2/6) GTCTC(1/5) G(A/G) GCATG G G(G/A/T)GC(C/A/T) GG G(A/G)GC(T/C) TT GACN G GACNNN TTT C AGC GT GACGT GAATGC(1/–1) C G ACTGGN TGG CC GAANN AT AT G CC AG T GT G(A/G) A CC G G T G G GGTAC GGGCC A A CCAN G GGTAC G T GCCNNNN GGATCNNNN CG(A/G)(T/C) GATNN CG(A/G)(T/C) CG(A/G)(T/C) CTGGAG(16/14) CAC GGCGC GGTGAN G (G/A) G GGATCNNNN G (G/A) t t t t t t t t t t t t t t t t t t t t t t t t t CCGGA CCGGA GATCA t t CCGGT CTAGT GATCT t t t t TTAAG TTAAG (C/T)CG(G/A)G TGCAC TAC G(C/T)(G/A)CC GTACC G(T/C)(A/G)CC CGCGC G(T/C)(A/G)CC GATCC G(A/T)CC GNCC GTACC TGCAC GGCCC t t CGAA TAAT TAAT AC (A/C)(G/T)AC t t t CT (C/G)GG TNAGG TNAGG t CGCC t t AAA CCA GTG 4 GCT t CCT AATT(C/T) AATT(C/T) t t t t t t t t t NNGTC t t CG(T/C)C CG(T/C)C NTGG C C C C NNATC 8 NNTTC t C C (1/–1) t NNGTC t NGGC t t t t t CG CG C t C t C EcoRII* BstOI BstNI BstEII BstBI Bst98I BssHII BsrSI BsrI BspEI BspDI BspCI Bsp1286I Bsp143II Bsp143I — Bsp120I Bsp119I Bsp106I Bsp68I Bsp19I BsoBI BsmAI BsmI Eco105I Eco91I Eco88I Eco81I Eco64I Eco52I Eco47III Eco47I Eco32I Eco24I EclXI EclHKI Ecl136II Eam1105I EagI DpnII* DraI DpnII DpnI DdeI CvnI Csp45I — Csp6I CspI CpoI ClaI Sau96I Cfr42I Hin6I, Cfr13I Cfr9I CfoI BsuRI Bsu36I Bsu15I EcoRII* BstZI BstYI BstXI 1

Eco52I, BanIII,

BstDEI 2

— CpoI,

Eco52I, – BstNI, 1 — BstPI, 1 BseNI,

AflII, BsePI, AhdI, Alw26I

CvnI, BstBI, AfeI

BmyI, BstOI Csp45I BsrSI AccIII ClaI PvuI HaeII MboI ApaI Csp45I ClaI NruI NcoI AvaI BsaMI SnaBI BstEII AvaI Bsu36I BanI BstZI AvaII EcoRV BanII BstZI SacI EcoICRI EclHKI DpnI MboI Bsu36I RsaI CspI SacII SmaI XmaI HhaI HaeIII ClaI XhoII ,

* , *, ,

, BfrI * , , ,

* BspDI AocI, ,

, , , Eam1105I, RsrII , , Ama87I,

RsrII SduI

,

Eco91I, BshNI,

HinP1I NspV, MvaI Bsp106I,

Sau3AI Sau3AI PauI

SinI , MflI XmaIII, EagI, BsrI FriOI AfaI* MvaI BseNI , ,

PalI BstZI EagI,

AhdI, NspV, NspV,

Eco81I Eco52I,

Bsp119I Eco64I XmaIII, , AspEI EagI, PspEI , , BcoI, Bsp119I BstBI EclXI, NdeII NdeII BspDI, AspEI cX C EclXI mII C XmaIII c8I C(C/T)CG(G/A)G Eco88I cX C EclXI

, mII C XmaIII

s1I AT Bsu15I ACTGGN(1/-1) GTCTC(1/5) G G G G TTT C G G(G/A/T)GC(C/A/T) TT AT TT AT GAATGCN AGC GACNNN GAG CCGC C TT GACNNN GT CG CG C AT CCANNNNN (A/G)GCGC C CC CC ACTGGN(1/-1) TCG CGAT CCC TAC GAT GG (A/G) CC GAGCT CC G(AG)GC(TC) CC GGGCC GA C T GCG G t t G t GA G G t G t t t t t t t t t t GATC CC(A/T)GG CC(A/T)GG GATC t t t me t t t t t t t t t t t t t CCGGA t t t t t t t t (TC)CG(AG)G GGCCG GGCCG GGCCG TNAG CCGGG GGCCG TTAAG CATGG t t GTNACC G(TC)(AG)CC G(A/T)CC TAC GNCC CGC GTNACC CGCGC GGCCC t A t CGAA CGAA CGAA t CGAT CGAT CGAT CGAT t t AC t t TC TC TNAGG TNAGG TNAGG (A/T)GG (A/T)GG t G(A/T)CCG G(A/T)CCG t CC t t AAA CTA t ATC CGA GCT GGG t CTC C TC GATC(T/C) CG t GG t C t t C t NNGTC NNGTC t t (T/C) t NTGG C t C PROMEGA CORPORATION 2800 WOODS HOLLOW ROAD MADISON, WI 53711-5399 USA TELEPHONE 608-274-4330 www.promega.com TECHNICAL REFERENCE

©2010 ALL RIGHTS RESERVED PART #GE633 Isoschizomers hI — EheI EcoT22I EcoT14I EcoRV — EcoRII EcoRI EcoICRI Eco147I Eco130I Enzyme (continued). Isoschizomers NspV NsiI NruI NotI NlaIII NheI NgoMIV NdeII BbeI* NdeI KasI* NcoI EheI* NciI NgoMIV EcoRII* NarI NaeI MvaI MstII MspA1I MspI MseI MscI MroI MluNI MluI MflI MboII MboI KspI — KpnI KasI I-PpoI Hsp92II Hsp92I HpaII HpaI HinP1I HinfI HindIII HinP1I*, HindII HincII Hin1I Eco47I, HhaI HgiEI HapII HaeIII HaeII FokI 2 BcnI

— — — Bsp19I EcoT22I, KspAI — — — Bsp68 I

3 Bsp143II

3

— AcyI,

1 —

Eco32I — BsuRI, HindII —

— NspBII AcyI,

Ecl136II NlaIII

­— ­— (s) Csp45I Hsp92II NaeI — MboI — BstOI Bsu36I HpaII Tru9I BalI AccIII BalI XhoII Sau3AI SacII Acc65I NarI MspI HhaI HincII CfoI HpaII NarI NsiI StyI BstOI SacI StuI StyI

, ,

* * *, * *, , BsaHI, Hsp92I ,

MluNI MscI

, , , *,

HapII Sau3AI PalI MspI HapII SstI* BstNI CfoI , *,

, SinI BstNI*,

Hin6I* BstBI, NdeII Mph1103I Asp718I* * , Hin1I

AvaII , , Bsp119I MvaI* DpnII DpnII

The enzymes in boldface type are available from Promega. from available are type boldface in enzymes The GAAGA(8/7) GGATG(9/13) G GCG C G(A/G) CATG ATGCA G(A/G) GG TT TCG GC GG C(A/C)G GG A GT(T/C) GCG GG C GAG C CATG ATGCA CC CC C T TGG TGG A (A/G) CCGC CC G C CTCTCTTAA C G GT(T/C) (A/G)GCGC GAGCT G GCC CC GGTAC GAT G T GTT AGG CA GGC t Sequence Recognition G t GGCGC G GGC G GCC t t G G G t t t t t t t t t t CC(A/T)GG GATC CC(A/T)GG GATC t t t t t t t t t t t t TAA CCGGA t AGCTT CGCGT t t t t CGG C(A/T)(A/T)GG C(A/T)(T/A)GG CATGG CGG CGG t ANTC CGC G(A/T)CC AATTC CTAGC CCGGC GCGCC CCGGC GTACC GCGCC CGC t t t t CGAA t t t t t TATG (A/T)GG (C/G)GG (A/T)GG TNAGG t t t t GGCCGC t t t t CC CGCC CGCC CGCC t AAC t t ATC CGA CCA CCA t CTC CCT GGC GGC C GCC GCC C GATC(T/C) t t t t t t GG t t t t CG(T/C)C CG(C/T)C T T C C C (A/G)AC (A/G)AC C(G/T)G t t (T/C) GGTAGC SinI SgfI SfuI SfiI SduI ScaI Sau96I Ecl136II*, Sau3AI SalI SacII SacI RsrII RsaI PvuII PvuI PstI PinAI PflMI PalI PaeR7I PaeI NspBII Enzyme Roberts, Reference Although 3. DpnI 2. 5 The 1. Notes: = * A, = N Key: XmnI XmaCI XmaIII XmaI XhoII XhoI ApaLI, XbaI VspI VneI Van91I TthHB8I Tth111I Tru9I TaqI StyI StuI SstII SstI SspI SphI SpeI SnaBI SmaI — — —

— applications. to methylation Sau3AI). cannot 3 of parentheses.

AclNI

EcoT14I

AatI, — TthHB8I SstI BspCI AseI, — AfaI — BstYI, PaeR7I —

′ MseI ...GTCTCN SstII,

Cfr9I, Asp700I Eco52I, a methylation Eco105I Cfr13I

locations AspI nucleotide

is R.J.

C, be unique

G

AvaII Csp45I Bsp1286I MboI CspI AgeI AccB7I HaeIII XhoI BbuI MspA1I Isoschizomer(s) SmaI XmaI SmaI AccB7I TaqI SacII EcoICRI SacI BbuI XmaI HpaII substituted — and ′ or ...CAGAGNNNNN t

of ...3 For T of

among

of either (adenine) and

,

, , Eco147I ′ Macelis, ,

KspI, cleavage , AsnI * * *,

XmaCI, CpoI example, MflI , SphI PaeI the NdeII Alw44I Eco47I

, , BsuRI

Cfr9I* BstZI

MspI *

Van91I PflMI

cytosine external for

EcoICRI commercially Cfr42I

, in recognize other sites , DpnII GTCTC(1/5) D. s its EagI,

...5

cytosine. recognition (1991) in falling enzymes ′ *

its EclXI

the available recognition outside indicates

Nucl. Acids Res. Acids Nucl. same These recognizing C sequence

restriction the nucleotide

enzymes

cleavage sequence, recognition in the order

enzymes

GATC may sequence, GAG G C C(A/C)G G(G/A/T)GC(C/A/T) CCGC CTGCA TT CG GCATG C GAGCT GT CGAT GAG G GACN T TAC C GAANN T AT C AAT C G C GGCCNNNN CAG GG AGT CCC CCC (A/G) CCGC CCC T T AGG GAGCT A CCAN GCGAT A CCAN G t Sequence Recognition GCATG at:

t t t t t t t t t t t t t GATC t t t t

while t 19 t CTAGA CGA TAA CGA t CTAGT CCGGT to

CCGGG GGCCG CCGGG TCGAG C(A/T)(A/T)GG CCGGG TCGAG t TGCAC G(A/T)CC GNCC TCGAC t site not CGAA t t TAAT t sequence AC t t t t t t t G(A/T)CCG CC t cut.

ATT GTA t ACT 4 4 CTG GGG GGG GGG t t t CCT CTC CTC (supp),

GATC(T/C) t t are be in MspI t CG t t t t t t NNGTC GG GG t NTGG NTGG

G C C CGC C C Therefore,

NNTTC requiring HpaII interchanged C(G/T)G indicated

is t (e.g.,

sensitive NGGCC 2077. is

sensitive methylation MboI DpnI in t C

for only and

to all