# Enzyme-Finder

---
url: https://www.nippongenetics.eu/en/enzyme-finder/
language: en
type: page
modified: 2022-09-27T09:21:54+02:00
---

     ![](https://www.nippongenetics.eu/app/uploads/dynamic/2021/02/keyvisual@3x-120x0-c-default.jpg) # Enzyme-Finder

Search for your restriction enzyme. Just add the name or a sequence to our enzyme finder.

[Back to products ‘Cloning’](https://www.nippongenetics.eu/en/product-category/cloning/)







 ## Search



 #### Ambiguity Codes

- n

     A or C or G or T (any)
- b

     C or G or T (not A)
- d

     A or G or T (not C)
- h

     A or C or T (not G)
- v

     A or C or G (not T)
- w

     A or T (weak)
- s

     C or G (strong)
- r

     A or G (purine)
- y

     C or T (pyrimidine)
- m

     A or C (amino)
- k

     G or T (keto)



      Name  Sequence ?  Overhang  Properties ?  Isoschizomere      [Aat II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/aat-ii/)   G↑ACGT↓C   3′ ACGT  - 37°
- IV
- 65°
- CpG
- FcB

   ZraI\*     [Acc I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/acc-i/)   GT↓MK↑AC   5′ MK  - 37°
- IV
- 80°
- CpG
- FcB

   Bsh1236I, BspFNI, BstFNI, BstUI, MvnI     [Acc III](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/acc-iii/)   T↓CCGG↑A   5′ CCGG  - 65°
- AccIII
- No
- Dam

   Aor13HI, BseAI, Bsp13I, BspEI, Kpn2I, MroI     [Acu I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/acu-i/)   CTGAAGN₁₄↑NN↓   3′ NN  - 37°
- IV
- 65°
- FcB

   Eco57I     [Afl II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/afl-ii/)   C↓TTAA↑G   5′ TTAA  - 37°
- IV
- 65°
- FcB

   BfrI, BspTI, BstAFI, MspCI, Vha464I     [Age I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/age-i/)   A↓CCGG↑T   5′ CCGG  - 37°
- IV
- 65°
- CpG
- FcB

   AsiGI, BshTI, CspAI, PinAI     [Alw I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/alw-i/)   GGATCNNNN↓N↑   5′ N  - 37°
- IV
- No
- Dam
- FcB

   AclWI, BspPI     [Alw26 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/alw26-i/)   GTCTCN↓NNNN↑   5′ NNNN  - 37°
- IV
- 65°
- CpG
- FcB

   BcoDI, BsmAI, BstMAI     [Apa I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/apa-i/)   G↑GGCC↓C   3′ GGCC  - 25°
- IV
- 65°
- CpG
- Dcm
- FcB

   Bsp120I\*, PspOMI\*     [ApaL I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/apal-i/)   G↓TGCA↑C   5′ TGCA  - 37°
- IV
- No
- CpG
- FcB

   Alw44I, VneI     [Apo I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/apo-i/)   R↓AATT↑Y   5′ AATT  - 50°
- III
- 80°
- FcB

   AcsI, XapI     [Asc I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/asc-i/)   GG↓CGCG↑CC   5′ CGCG  - 37°
- IV
- 65°
- CpG
- FcB

   PalAI, SgsI     [Ava I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ava-i/)   C↓YCGR↑G   5′ YCGR  - 37°
- IV
- 80°
- CpG
- FcB

   Ama87I, BmeT110I, BsiHKCI, BsoBI, Eco88I     [Ava II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ava-ii/)   G↓GWC↑C   5′ GWC  - 37°
- IV
- 80°
- CpG
- Dcm
- FcB

   Bme18I, Eco47I, SinI, VpaK11BI     [Avr II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/avr-ii/)   C↓CTAG↑G   5′ CTAG  - 37°
- IV
- 80°
- FcB

   AspA2I, BlnI, XmaJI     [Bal I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bal-i/)   TGG⇅CCA   Blunt  - 37°
- BalI
- 65°
- Dcm

   MlsI, MluNI, Mox20I, MscI, Msp20I     [BamH I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bamh-i/)   G↓GATC↑C   5′ GATC  - 37°
- BamHI
- No
- FcB

   -     [Bcl I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bcl-i/)   T↓GATC↑A   5′ GATC  - 50°
- III
- No
- Dam
- FcB

   FbaI, Ksp22I     [Bgl I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bgl-i/)   GCCN↑NNN↓NGGC   3′ NNN  - 37°
- III
- 65°
- CpG
- FcB

   -     [Bgl II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bgl-ii/)   A↓GATC↑T   5′ GATC  - 37°
- III
- No
- FcB

   -     [Bsa I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bsa-i/)   GGTCTCN↓NNNN↑   5′ NNNN  - 37°
- IV
- 65°
- CpG
- Dcm
- FcB

   Bso31I, BspTNI, Eco31I     [BsaW I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bsaw-i/)   W↓CCGG↑W   5′ CCGG  - 60°
- IV
- 80°
- FcB

   -     [BsiW I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bsiw-i/)   C↓GTAC↑G   5′ GTAC  - 55°
- III
- 80°
- CpG
- FcB

   Pfl23II, PspLI     [BsmB I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bsmb-i/)   CGTCTCN↓NNNN↑   5′ NNNN  - 55°
- III
- 80°
- CpG
- FcB

   Esp3I     [BsoB I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bsob-i/)   C↓YCGR↑G   5′ YCGR  - 37°
- IV
- 80°
- FcB

   Ama87I, AvaI, BmeT110I, BsiHKCI, Eco88I     [BspE I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bspe-i/)   T↓CCGG↑A   5′ CCGG  - 37°
- III
- 80°
- CpG
- Dam
- FcB

   AccIII, Aor13HI, BseAI, Bsp13I, Kpn2I, MroI     [BsrF I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bsrf-i/)   R↓CCGG↑Y   5′ CCGG  - 37°
- IV
- No
- CpG
- FcB

   Bse118I, BssAI, Cfr10I     [BstY I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/bsty-i/)   R↓GATC↑Y   5′ GATC  - 60°
- II
- 80°
- FcB

   BstX2I, MflI, PsuI     [BtsC I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/btsc-i/)   GGATG↑NN↓   3′ NN  - 50°
- IV
- 80°
- FcB

   BseGI, BstF5I, FokI\*     [Cfr10 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/cfr10-i/)   R↓CCGG↑Y   5′ CCGG  - 37°
- Cfr10I
- No
- CpG
- FcB

   Bse118I, BsrFI, BssAI     [Cfr42 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/cfr42-i/)   CC↑GC↓GG   3′ GC  - 37°
- I
- 65°
- CpG
- FcB

   KspI, SacII, Sfr303I, SgrBI     [Cfr9 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/cfr9-i/)   C↓CCGG↑G   5′ CCGG  - 37°
- III
- 65°
- CpG

   SmaI\*, TspMI, XmaI     [Cla I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/cla-i/)   AT↓CG↑AT   5′ CG  - 37°
- IV
- 65°
- CpG
- Dam
- FcB

   Bsa29I, BseCI, BshVI, BspDI, Bsu15I, BsuTUI     [CviA I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/cvia-i/)   ↓GATC↑   5′ GATC  - 37°
- IV
- 65°
- Dam
- FcB

   -     [Dde I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/dde-i/)   C↓TNA↑G   5′ TNA  - 37°
- III
- 65°
- FcB

   BstDEI, HpyF3I     [Dpn I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/dpn-i/)   GA⇅TC   Blunt  - 37°
- IV
- 80°
- FcB

   MalI     [Dpn II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/dpn-ii/)   ↓GATC↑   5′ GATC  - 37°
- DpnII
- 65°
- Dam
- FcB

   Bsp143I, BssMI, BstKTI\*, BstMBI, Kzo9I, MboI, NdeII, Sau3AI     [Dra I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/dra-i/)   TTT⇅AAA   Blunt  - 37°
- IV
- 65°
- FcB

   -     [Eag I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/eag-i/)   C↓GGCC↑G   5′ GGCC  - 37°
- III
- 65°
- CpG
- FcB

   BseX3I, BstZI, EclXI, Eco52I     [Eco47 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/eco47-i/)   G↓GWC↑C   5′ GWC  - 37°
- III
- 65°
- CpG
- Dcm
- FcB

   AvaII, Bme18I, SinI, VpaK11BI     [EcoN I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/econ-i/)   CCTNN↓N↑NNAGG   5′ N  - 37°
- IV
- 65°
- FcB

   BstENI, XagI     [EcoO109 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ecoo109-i/)   RG↓GNC↑CY   5′ GNC  - 37°
- IV
- 65°
- Dcm
- FcB

   -     [EcoR I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ecor-i/)   G↓AATT↑C   5′ AATT  - 37°
- EcoRI
- 65°
- CpG
- FcB

   -     [EcoR V](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ecor-v/)   GAT⇅ATC   Blunt  - 37°
- III
- 65°
- CpG
- FcB

   Eco32I     [EcoT38 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ecot38-i/)   G↑RGCY↓C   3′ RGCY  - 37°
- IV
- 65°
- FcB

   BanII, Eco24I, FriOI     [Esp3 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/esp3-i/)   CGTCTCN↓NNNN↑   5′ NNNN  - 37°
- IV
- 65°
- CpG
- FcB

   BsmBI     [Fok I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/fok-i/)   GGATGN₉↓NNNN↑   5′ NNNN  - 37°
- IV
- 65°
- CpG
- Dcm
- FcB

   BseGI\*, BstF5I\*, BtsCI\*     [Fsp I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/fsp-i/)   TGC⇅GCA   Blunt  - 37°
- IV
- 65°
- CpG
- FcB

   Acc16I, NsbI     [Hae II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hae-ii/)   R↑GCGC↓Y   3′ GCGC  - 37°
- IV
- 80°
- CpG
- FcB

   BfoI, BstH2I     [Hae III](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hae-iii/)   GG⇅CC   Blunt  - 37°
- IV
- 80°
- FcB

   BshFI, BsnI, BimgI, BsuRI     [Hga I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hga-i/)   GACGCN₅↓NNNNN↑   5′ NNNNN  - 37°
- I
- 65°
- CpG
- FcB

   CseI     [Hinc II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hinc-ii/)   GTY⇅RAC   Blunt  - 37°
- IV
- 65°
- CpG
- FcB

   HindII     [XmaI](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/xmai/)   C↓CCGG↑G   5′ CCGG  - 37°
- IV
- 65°
- CpG
- FcB

   Cfr9I, SmaI\*, TspMI     [Hind II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hind-ii/)   GTY⇅RAC   Blunt  - 37°
- II
- 65°
- CpG
- FcB

   HincII     [Hind III](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hind-iii/)   A↓AGCT↑T   5′ AGCT  - 37°
- II
- 80°
- FcB

   -     [Hinf I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hinf-i/)   G↓ANT↑C   5′ ANT  - 37°
- IV
- 80°
- CpG
- FcB

   -     [HinP1 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hinp1-i/)   G↓CG↑C   5′ CG  - 37°
- II
- 65°
- CpG
- FcB

   AspLEI\*, BstHHI\*, CfoI\*, HhaI\*, Hin6I, HspAI     [Hpa I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hpa-i/)   GTT⇅AAC   Blunt  - 37°
- IV
- No
- CpG
- FcB

   KspAI     [Hpa II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hpa-ii/)   C↓CG↑G   5′ CG  - 37°
- IV
- 80°
- CpG
- FcB

   BsiSI, HapII, MspI     [Hph I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hph-i/)   GGTGAN₇↑N↓   3′ N  - 37°
- IV
- 65°
- Dam
- FcB

   AsuHPI     [Hpy188 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hpy188-i/)   TC↑N↓GA   3′ N  - 37°
- IV
- 65°
- Dam
- FcB

   -     [Hpy99 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hpy99-i/)   ↑CGWCG↓   3′ CGWCG  - 37°
- IV
- 65°
- CpG
- FcB

   -     [HpyCH4 V](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/hpych4-v/)   TG⇅CA   Blunt  - 37°
- IV
- 65°
- FcB

   -     [Kpn I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/kpn-i/)   G↑GTAC↓C   3′ GTAC  - 37°
- I
- No
- FcB

   Acc65I\*, Asp718I\*     [Kpn2 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/kpn2-i/)   T↓CCGG↑A   5′ CCGG  - 55°
- I
- 80°
- CpG
- FcB

   AccIII, Aor13HI, BseAI, Bsp13I, BspEI, MroI     [Lsp1109 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/lsp1109-i/)   GCAGCN₈↓NNNN↑   5′ NNNN  - 37°
- III
- 65°
- FcB

   BbvI, BseXI, BstV1I     [Mbo I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/mbo-i/)   ↓GATC↑   5′ GATC  - 37°
- III
- 65°
- CpG
- Dam
- FcB

   Bsp143I, BssMI, BstKTI\*, BstMBI, DpnII, Kzo9I, NdeII, Sau3AI     [Mbo II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/mbo-ii/)   GAAGAN₇↑N↓   3′ N  - 37°
- II
- 65°
- Dam
- FcB

   -     [Mlu I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/mlu-i/)   A↓CGCG↑T   5′ CGCG  - 37°
- III
- 65°
- CpG
- FcB

   -     [Xho I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/xho-i/)   C↓TCGA↑G   5′ TCGA  - 37°
- IV
- 80°
- CpG
- FcB

   PaeR7I, Sfr274I, SlaI     [Mnl I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/mnl-i/)   CCTCN₆↑N↓   3′ N  - 37°
- II
- 65°
- FcB

   -     [Mse I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/mse-i/)   T↓TA↑A   5′ TA  - 37°
- IV
- 65°
- FcB

   SaqAI, Tru1I, Tru9I     [Msp I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/msp-i/)   C↓CG↑G   5′ CG  - 37°
- IV
- No
- FcB

   BsiSI, HapII, HpaII     [MspA1 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/mspa1-i/)   CMG⇅CKG   Blunt  - 37°
- IV
- 65°
- CpG
- FcB

   -     [Mun I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/mun-i/)   C↓AATT↑G   5′ AATT  - 37°
- II
- 65°
- FcB

   MfeI     [Nae I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/nae-i/)   GCC⇅GGC   Blunt  - 37°
- I
- 65°
- CpG
- FcB

   MroNI\*, NgoMIV\*, PdiI     [Nco I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/nco-i/)   C↓CATG↑G   5′ CATG  - 37°
- III
- 65°
- FcB

   Bsp19I     [Nde I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/nde-i/)   CA↓TA↑TG   5′ TA  - 37°
- IV
- 65°
- FcB

   FauNDI     [NgoM IV](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ngom-iv/)   G↓CCGG↑C   5′ CCGG  - 37°
- IV
- 80°
- CpG
- FcB

   MroNI, NaeI\*, PdiI\*     [Nhe I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/nhe-i/)   G↓CTAG↑C   5′ CTAG  - 37°
- II
- 65°
- CpG
- FcB

   AsuNHI, BmtI\*, BspOI\*     [Nla IV](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/nla-iv/)   GGN⇅NCC   Blunt  - 37°
- IV
- 65°
- CpG
- Dcm
- FcB

   BmiI, BspLI, PspN4I     [Not I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/not-i/)   GC↓GGCC↑GC   5′ GGCC  - 37°
- III
- 65°
- CpG
- FcB

   CciNI     [Nru I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/nru-i/)   TCG⇅CGA   Blunt  - 37°
- III
- 65°
- CpG
- Dam
- FcB

   Bsp68I, BtuMI, RruI     [Nt.BstNB I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/nt-bstnb-i/)   GAGTCNNNN↓   Nicht vorhanden  - 55°
- III
- 80°
- FcB

   -     [PaeR7 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/paer7-i/)   C↓TCGA↑G   5′ TCGA  - 37°
- IV
- No
- CpG
- FcB

   Sfr274I, SlaI, XhoI     [PflM I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/pflm-i/)   CCAN↑NNN↓NTGG   3′ NNN  - 37°
- III
- 65°
- Dcm
- FcB

   AccB7I, Van91I     [Ple I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ple-i/)   GAGTCNNNN↓N↑   5′ N  - 37°
- IV
- 65°
- CpG
- FcB

   MlyI\*, PpsI, SchI\*     [PluT I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/plut-i/)   G↑GCGC↓C   3′ GCGC  - 37°
- IV
- 65°
- CpG
- FcB

   DinI\*, EgeI\*, EheI\*, KasI, SfoI\*     [PspG I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/pspg-i/)   ↓CCWGG↑   5′ CCWGG  - 37°
- IV
- No
- Dcm

   AjnI, BciT130I\*, BseBI\*, BstNI\*, Bst2UI\*, EcoRII, MvaI\*, Psp6I     [Pst I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/pst-i/)   C↑TGCA↓G   3′ TGCA  - 37°
- III
- 80°
- FcB

   BspMAI     [Xba I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/xba-i/)   T↓CTAG↑A   5′ CTAG  - 37°
- IV
- 65°
- Dam
- FcB

   -     [Tth111 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/tth111-i/)   GACN↓N↑NGTC   5′ N  - 65°
- IV
- No
- FcB

   PflFI, PsyI     [TspM I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/tspm-i/)   C↓CCGG↑G   5′ CCGG  - 75°
- IV
- No
- CpG
- FcB

   Cfr9I, SmaI\*, XmaI     [Taq I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/taq-i/)   T↓CG↑A   5′ CG  - 65°
- III
- 80°
- Dam
- FcB

   -     [Swa I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/swa-i/)   ATTT⇅AAAT   Blunt  - 25°
- III
- 65°
- FcB

   SmiI     [Pvu I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/pvu-i/)   CG↑AT↓CG   3′ AT  - 37°
- III
- No
- CpG
- FcB

   Ple19I     [Pvu II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/pvu-ii/)   CAG⇅CTG   Blunt  - 37°
- II
- No
- FcB

   -     [Rsa I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/rsa-i/)   GT⇅AC   Blunt  - 37°
- IV
- 65°
- CpG
- FcB

   AfaI, Csp6I\*, CviQI\*, RsaNI\*     [Sac I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sac-i/)   G↑AGCT↓C   3′ AGCT  - 37°
- I
- 65°
- FcB

   Ecl136II\*, EcoICRI\*, Eco53kI\*, Psp124BI, SstI     [Sac II](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sac-ii/)   CC↑GC↓GG   3′ GC  - 37°
- IV
- 65°
- CpG
- FcB

   Cfr42I, KspI, Sfr303I, SgrBI     [Sal I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sal-i/)   G↓TCGA↑C   5′ TCGA  - 37°
- III
- 65°
- CpG
- FcB

   -     [Sau96 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sau96-i/)   G↓GNC↑C   5′ GNC  - 37°
- IV
- 80°
- CpG
- Dcm
- FcB

   AspS9I, BmgT120I, Cfr13I, PspPI     [Sbf I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sbf-i/)   CC↑TGCA↓GG   3′ TGCA  - 37°
- IV
- 80°
- FcB

   SdaI, Sse8387I     [Sca I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sca-i/)   AGT⇅ACT   Blunt  - 37°
- III
- 80°
- FcB

   ZrmI     [Sda I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sda-i/)   CC↑TGCA↓GG   3′ TGCA  - 37°
- IV
- 80°
- FcB

   SbfI, Sse8387I     [Sfi I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sfi-i/)   GGCCN↑NNN↓NGGCC   3′ NNN  - 50°
- II
- No
- CpG
- Dcm
- FcB

   -     [SgrA I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sgra-i/)   CR↓CCGG↑YG   5′ CCGG  - 37°
- IV
- 65°
- CpG
- FcB

   -     [Sma I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sma-i/)   CCC⇅GGG   Blunt  - 25°
- IV
- 65°
- CpG
- FcB

   Cfr9I\*, TspMI\*, XmaI\*     [SnaB I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/snab-i/)   TAC⇅GTA   Blunt  - 37°
- IV
- 80°
- CpG
- FcB

   BstSNI, Eco105I     [Spe I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/spe-i/)   A↓CTAG↑T   5′ CTAG  - 37°
- IV
- 80°
- FcB

   AhlI, BcuI     [Sph I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sph-i/)   G↑CATG↓C   3′ CATG  - 37°
- II
- 65°
- FcB

   PaeI     [Sse9 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/sse9-i/)   ↓AATT↑   5′ AATT  - 55°
- I
- 65°
- FcB

   MluCI, TasI     [Ssp I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/ssp-i/)   AAT⇅ATT   Blunt  - 37°
- IV
- 65°
- FcB

   -     [Stu I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/stu-i/)   AGG⇅CCT   Blunt  - 37°
- IV
- 65°
- Dcm
- FcB

   Eco147I, PceI, SseBI     [StyD4 I](https://www.nippongenetics.eu/en/products/cloning/restriction-enyzme-finder/styd4-i/)   ↓CCNGG↑   5′ CCNGG  - 37°
- IV
- 65°
- CpG
- Dcm
- FcB

   Bme1390I\*, BmrFI\*, BstSCI, MspR9I\*, ScrFI\*













×













×

- 25°
- 37°
- 50°
- 55°
- 60°
- 65°
- 75°

- AccIII
- BalI
- BamHI
- Cfr10I
- DpnII
- EcoRI
- I
- II
- III
- IV

- 65°
- 80°
- No

- CpG
- Dam
- Dcm

- FcB















     ![](https://www.nippongenetics.eu/app/uploads/2021/02/keyvisual@3x-120x0-c-default.jpg) ## Double digest − Buffer compatibility chart

Performing two digestions simultaneously saves a lot of time.

Check the buffer compatibility at the FastGene® Double Digest Chart:

[DOWNLOAD](https://www.nippongenetics.eu/app/uploads/2021/05/restriction-enzymes-double-digest-symbols.xlsx)





 ![](https://www.nippongenetics.eu/app/uploads/2021/05/double-digest-buffer-compatibility-120x0-c-default.jpg)







## Restriction Endonucleases

**What are Restriction Enzymes?**

Restriction enzmyes recognize short DNA sequences and cleaves double-stranded DNA at or near a specific recognition site. Restriction enzmyes are classified into four types, based on their subunit structure, cofactor requirements and specificity of cleavage.

3,000 different restriction enzymes have been discovered, which recognize over 230 distinct DNA sequences. These enzymes are routinely used for DNA modification around the world and are an indispensable tool in molecular cloning.

**Historical background**

The basis for the research of restriction enzymes goes back to the work of Luria and colleagues in the early 1950s \[1\]. Luria observed that the bacteriophage λ can grow good in one strain of _E.coli (e.g. E.coli C)_, but often poorly in another _E.coli_ strain _(e.g. E.coli_ _K_). The host cell (_E.coli_ _K)_ was known as the restriction host and appears to have the ability to reduce the biological activity of the phage λ.

The first-time that the term restriction enzyme was mentioned was in the 1960s in the laboratories of Arber and Meselson. They found out that the restriction is caused by an enzymatic cleavage of the phage DNA. The enzyme involved in this process was termed “restriction enzyme” \[2, 3\]. The restriction enzymes studied by Arber and Meselson were type I restriction enzymes, which cleave DNA at random places away from the recognition site.

In 1970, Smith and colleagues isolated and describe the first type II restriction enzyme, Hind II \[4\]. Restriction enzymes of type II are much more useful for laboratory work, because they cleave DNA at the site of their recognition sequence. Due to its importance for molecular biology, Smith, Arber and Nathans shared the 1978 Nobel Prize for Medicine and Physiology for their discovery of restriction enzymes and their application to molecular genetics.

**Recognition sequences**

All restriction endonucleases recognize a specific DNA sequence. The recognition sequence is usually palindromic or partially palindromic, meaning the base sequence reads the same backwards and forwards. Restriction enzymes can cleave double stranded DNA either at the center of both strands to yield “blunt ends” or at a staggered position leaving overhangs called “sticky ends”.

**Different Types of restriction enzymes**

Based on the structure, cofactor requirements and specificity of cleavage there are four types of restriction enzymes (Types I, II, III, and IV).

**Type I** restriction enzymes cleaves the DNA at a random location far away from the recognition sequence. These enzymes require both ATP and S-adenosyl-L-methionine to function.

**Type II** restriction enzymes cleaves the DNA within or near the recognition sequence. These enzymes do not require ATP and are independent from methylase. The type II enzymes are the most useful restriction endonucleases for the daily laboratory work. All our NIPPON Genetics EUROPE restriction enzymes are from type II.

**Type III** restriction enzymes cleaves DNA about 20 – 25 base pairs away from the recognition sequence. They require both ATP and S-adenosyl-L-methionine to function.

**Type IV** restriction enzymes cleaves only modified, typically methylated DNA in contrast to the types I-III, which are usually inhibited by methylation.

**Double digestion**

A vector and an insert DNA can be cloned by cleaving with two different restriction enzymes, thus generating two different restriction ends. This strategy prevents the vector from being ligated without an insert, resulting in great reduction in self-ligation and increase in cloning efficiency. Most of our restriction enzymes are 100% active in the FastCut Buffer, making double digestion simple. Please look at the **Double Digest Chart** to make a double digestion with the four standard buffers.

[DOUBLE DIGEST CHART (DOWNLOAD)](https://www.nippongenetics.eu/app/uploads/2021/05/restriction-enzymes-double-digest-symbols.xlsx)

**Activity of Unique FastGene® Buffer**

Nippon Genetics provides four standard buffers that maximally support the activity of each restriction enzyme in the buffer provided with the enzyme. However, some restriction endonucleases require unique buffer for maximal activity. Take a look on the **Double Digest Chart** to select a buffer for double digestion if a restriction enzyme requires unique buffer. Listed are activities (%) of the most commonly used restriction endonucleases in the five unique FastGene® Buffers for EcoR I, BamH l, Acc III, Bal I, and Dpn Il, respectively. A restriction enzyme, if it is active in one of the four standard buffers, is usually active in a unique buffer. Therefore, it is possible to perform double digestion in a particular unique buffer. If digestion efficiency is low due to the suboptimal buffer, increase the amount of restriction endonucleases or incubate for a longer period of time.

[DOUBLE DIGEST CHART (DOWNLOAD)](https://www.nippongenetics.eu/app/uploads/2021/05/restriction-enzymes-double-digest-symbols.xlsx)

##### **References**

\[1\] Luria and Human (1952) A nonhereditary, host-induced variation of bacterial viruses. _J Bacteriol_., 557-569.

\[2\] Arber and Linn (1969) DNA modification and restriction. _Annu Rev Biochem_., 467-500.

\[3\] Meselson and Yuan (1968) DNA restriction enzyme from E. coli. _Nature_, 1110-1114

\[4\] Smith and Wilcox (1970) A restriction enzyme from Hemophilus influenzae. I. Purification and general properties. _J Mol Biol_., 379-391.