Restriction Enzymes

Restriction enzymes cut DNA at a specific sequence, making them very useful in a variety of applications, including molecular cloning, SNP analysis, Restriction Fragment Length Polymorphism (RFLP), and preparation of DNA for Southern blots.
 

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Restriction Enzyme

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APAI-RO Apa I from Acetobacter pasteurianus Recognition sites: GGG*CC*C
GGG*CC*C
Restriction site: GGG*CC↓*C
GGG*CC↓*C
Heat inactivation: Apa I can be heat inactivated by incubation at 65 °C for 15 minutes.
BLNI-RO Bln I (Avr II) from Brevibacterium linens Recognition sites: CCTAGG
CCTAGG
Restriction site: C↓CTAGG
C↓CTAGG
Heat inactivation: No inactivation of Bln I after incubation at 65 °C for 15 minutes.
DPNI-RO Dpn I from Diplococcus pneumoniae Recognition sites: GmAT*C
GmAT*C
Restriction site: GmA↓T*C
GmA↓T*C
Heat inactivation: No inactivation of Dpn I after incubation at 65 °C for 15 minutes.
R3884 EclX I from Enterobacter cloacae 590 Restriction Enzyme Recognition sequence: 5′-C/GGCCG-3′
Ligation and recutting results: After 2-10-fold EclX I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >95% of fragments can be ligated, and >95% recut.
Heat inactivation: 65 °C for 20 minutes.
MAEIII-RO Mae III from Methanococcus aeolicus PL-15/H Recognition sites: GTNAC
GTNAC
Restriction site: ↓GTNAC
↓GTNAC
Heat inactivation: There is no information available whether or not Mae III can be heat inactivated.
NCOI-RO Nco I from Rhodococcusspecies (formerly Nocardia corallina) Nco I recognizes the sequence *C↓C°ATGG and generates fragments with 5′-cohesive termini.
Recognition sites: *CC°ATGG
*CC°ATGG
Restriction site: *C↓C°ATGG
*C↓C°ATGG
Heat inactivation: Nco I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
NHEI-RO Nhe I from Neisseria mucosa Recognition sites: GCTAG*C
GCTAG*C
Restriction site: G↓CTAG*C
G↓CTAG*C
Heat inactivation: Nhe I can be heat inactivated by incubation at 65 °C for 15 minutes.
Star activity: Use of high concentrations of Nhe I leads to star activity; the additional bands which occur in the case of star activity might give the impression of partial cutting (if a defined pattern is expected).
Methylation sensitivity: Nhe I is inhibited by the presence of 5′-methylcytosine at the site indicated (*) in the sequence G/CTAG*C.
NOTI-RO Not I from Nocardia otitidis-caviarum Recognition sites: GCGG*C*CG °C
GCGG*C*CG °C
Restriction site: GC↓GG*C*CG °C
GC↓GG*C*CG °C
Heat inactivation: Not I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
PVUI-RO Pvu I from Proteus vulgaris Pvu I recognizes the sequence CG°AT↓ *CG and generates fragments with 3′-cohesive termini.
Recognition sites: CG°AT*CG
CG°AT*CG
Restriction site: CG°AT↓*CG
CG°AT↓*CG
Heat inactivation: No inactivation of Pvu I after incubation at 65 °C for 15 minutes.
SACI-RO Sac I (Sst I) from Streptomyces achromogenes Recognition sites: G°AG*CTC
G°AG*CTC
Restriction site: G°AG*CT↓C
G°AG*CT↓C
Heat inactivation: Sac I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
SFII-RO Sfi I from Streptomyces fimbriatus Recognition sites: GG °C*CN5GG °C °C
GG °C*CN5GG °C °CGG °C*CNNNNNGG °C °C
Restriction site: GG °C*CN4↓NGG °C °C
GG °C*CN4↓NGG °C °CGG °C*CNNNN↓NGG °C °C
Heat inactivation: No inactivation of Sfi I after incubation at 65 °C for 15 minutes.
Compatible ends
For an Sfi I end to be compatible and ligatable with any other end, the two corresponding recognition sites must have contained the same three internal Ns.
Isoschizomers
The enzyme has no known isoschizomers.
Methylation sensitivity
Sfi I is inhibited by the presence of 5-methylcytosine at the most central C residue (*) on the recognition sequence. 5-methylcytosine at the other C-positions (°) will not inhibit the enzyme.
Incubation temperature
+50°C
SMAI-RO Sma I from Serratia marcescens Sb Recognition sites: *C °C*CGGG
*C °C*CGGG
Restriction site: *C °C*C↓GGG
*C °C*C↓GGG
Heat inactivation: Sma I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
SPEI-RO Spe I from Sphaerotilus species Recognition sites: *A*CTAGT
*A*CTAGT
Restriction site: *A↓*CTAGT
*A↓*CTAGT
Heat inactivation: Spe I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
11371517001 Swa I from Staphylococcus warneri Recognition sites: ATTTAAAT
ATTTAAAT
Restriction site: ATTT↓AAAT
ATTT↓AAAT
Heat inactivation: Swa I can be heat inactivated by incubation at 65 °C for 20 minutes.
TAQI-RO Taq I from Thermus aquaticus YTI Recognition sites: TCG*A
TCG*A
Restriction site: T↓CG*A
T↓CG*A
Heat inactivation: No inactivation of Taq I after incubation at 65 °C for 15 minutes.
XBAI-RO Xba I from Xanthomonas campestris Star Activity
The sequence specificity of Xba I is relaxed at low ionic strength or by addition of glycerol, ethanol or DMSO to the incubation mixture.
Recognition sites: TCTAGA
TCTAGA
Restriction site: T↓CTAGA
T↓CTAGA
Heat inactivation: Xba I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 15 U/μg DNA). Higher concentrations of Xba I cannot be heat inactivated completely under these conditions.