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.
 

Product #

Restriction Enzyme

Description

Add to Cart

R6142 Acc I from Acinetobacter calcoaceticus Restriction Enzyme Recognition Sequence: 5′-GT/(A,C)(G,T)AC-3′
Cutting results: a 2-10-fold Acc I overdigestion of 1 μg pBR322 DNA substrate results in 100% cutting
Heat inactivation: Activity retained after 10 minutes at 65 °C. Five-fold enhanced activity occurs at 55°C. Enzyme inactivated at 80 °C for 20 minutes.
11209183001 Afl III from Anabaena flos-aquae Recognition sites: A*CRYGT
A*CRYGTA*C(A/G)(T/C)GT
Restriction site: A↓*CRYGT
A↓*CRYGTA↓*C(A/G)(T/C)GT
Heat inactivation: Afl III is not heat-inactivated by heating to 65 °C for 20 min.
50% acitvity remain after 20 minutes incubation at 65 °C.
R6885 Alu I from Arthrobacter luteus Restriction Enzyme Recognition Sequence: 5′-AG/CT-3′
Ligation and recutting results: After 2-10-fold Alu I overdigestion of 1 μg λ DNA substrate, results in 95% cutting, >80% of fragments can be ligated, and >95% recut.
Heat inactivation: Inactivated at 65 °C for 15 minutes.
ALUI-RO Alu I from Arthrobacter luteus Recognition sites: *AG*CT
*AG*CT
Restriction site: *AG↓*CT
*AG↓*CT
Heat inactivation: Alu I can be heat inactivated by incubation at 65 °C for 15 minutes.
R4258 Apa I from Acetobacter pasteurianus Restriction Enzyme Recognition Sequence: 5′-GGGCC/C-3′
Cutting results: a 2-10-fold Apa I overdigestion of Ad-2 DNA substrate results in 100% cutting.
Heat inactivation: 65 °C for 15 minutes.
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.
ASP718-RO Asp718 I from Acinetobacter species 718 Recognition sites: GGT°AC*C
GGT°AC*C
Restriction site: G↓GT°AC*C
G↓GT°AC*C
Heat inactivation: There is no information available whether or not Asp718 I can be heat inactivated.
R0260 BamH I from Bacillus amyloliquefaciens H Restriction Enzyme Recognition sequence: 5′-G/GATCC-3′
Cutting results: a 2-10-fold Bam HI overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: 60 °C for 15 minutes.
Star activity: To prevent star activity, avoid suboptimal reaction conditions containing low salt concentration, high glycerol (>5%) and high pH 8.0.
BAMHI-RO BamH I from Bacillus amyloliquefaciens (H) Specificity: Star Activity
BamH I exhibits star activity under non-optimal conditions. The relaxed specificities of BamH I are GGATCN or G(R)ATCC. The relaxed specificity of BamH I can be induced by lowering the ionic strength, by glycerol concentration higher than 5% or by using excess enzyme.
Recognition sites: GG°AT*C °C
GG°AT*C °C
Restriction site: G↓G°AT*C °C
G↓G°AT*C °C
Methylation sensitivity: BamH I is not inhibited by overlapping dam-methylation, but is inhibited by the presence of 5- or 4-methylcytosine at the internal C residue indicated (*) in the recognition sequence.
Heat inactivation: BamH I can be heat inactivated by incubation at 65 °C for 15 minutes (tested with up to 10 U/μg DNA). If higher enzyme concentrations are used, BamH I is resistant to heat inactivation.
BCLI-RO Bcl I Recognition sites: TG*AT*CA
TG*AT*CA
Restriction site: T↓G*AT*CA
T↓G*AT*CA
Heat inactivation: No inactivation of Bcl I after incubation at 65 °C for 15 minutes.
R8631 Bcl I from Bacillus caldolyticus Restriction Enzyme Recognition sequence: 5′-T/GATCA-3′
Cutting results: a 2-10-fold Bcl I overdigestion of 1 μg λ DNA substrate results in 100% cutting.
Heat inactivation: This enzyme cannot be heat inactivated at 65 °C for 15 minutes.
11198939001 Bfr I (Afl II) from Bacteroides fragilis Recognition sites: *CTTAAG
*CTTAAG
Restriction site: *C↓TTAAG
*C↑TTAAG
Heat inactivation: Bfr I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
R6377 Bgl II from Bacillus licheniformis Restriction Enzyme Recognition sequence: 5′-A/GATCT-3′
Cutting results: a 2-10-fold Bgl II overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat Inactivation: This enzyme cannot be heat inactivated.
BGLII-RO Bgl II from Bacillus globigii Recognition sites: AG°AT*CT
AG°AT*CT
Restriction site: A↓G°AT*CT
A↓G°AT*CT
Heat inactivation: No inactivation of Bgl II after 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.
R3131 Bln I from Brevibacterium linens buffered aqueous glycerol solution Recognition sequence: 5′-C/CTAGG-3′
Ligation and recutting results: After 2-10-fold Bln I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >70% of fragments can be ligated, and >90% recut.
Heat inactivation:. This enzyme is not heat inactivated at 65 °C for 15 minutes.
R3635 Bsm I from Bacillus stearothermophilus NUB 36 Restriction Enzyme Recognition sequences: 5′-GAATGCN/-3′ 3′-CTTAC/GN 5
Ligation and recruiting results: After 2-10-fold Bsm I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >90% of fragments can be ligated, and >95% recut.
Heat inactivation: Heat inactivation for Bsm I is not available.
11292307001 Bsm I from Bacillus stearothermophilus NUB36 Recognition sites: G*AATG °CNN
G*AATG °CNN
Restriction site: G*AATG °CN↓N
G*AATG °CN↓NGAATGC (1/-1)
Heat inactivation: There is no information available whether or not Bsm I can be heat inactivated.
R4253 BstE II from Bacillus stearothermophilus ET Restriction Enzyme Recognition sequence: 5′-G/GTNACC-3′
Cutting results: a 2-10-fold BstE II overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: This enzyme cannot be inactivated at 65 °C for 15 minutes.
10688541001 Cfo I (Hha I) from Clostridium formicoaceticum Recognition sites: G*CG*C
G*CG*C
Restriction site: G*CG↓*C
G*CG↓*C
Heat inactivation: There is no information available whether or not Cfo I can be heat inactivated.
R1761 Cfo I from Clostridium formicoaceticum Restriction Enzyme Recognition Sequence: 5′-GCG/C-3′
Cutting results: a 2-10-fold Cfo I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Not complete at 65 °C for 10 minutes.
R7763 Cla I from Caryophanon latum L Restriction Enzyme Recognition sequence: 5′-AT/CGAT-3′
Cutting results: a 2-10-fold Cla I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Enzyme is not inactivated at 65 °C for 20 minutes.
CLAI-RO Cla I from Caryophanon latum L Recognition sites: *AT*CG*AT
*AT*CG*AT
Restriction site: *AT↓*CG*AT
*AT↓*CG*AT
Heat inactivation: No inactivation of Cla I after incubation at 65 °C for 15 minutes.
R4256 Dde I from Desulfovibrio desulfuricans strain Norway Restriction Enzyme Recognition sequence: 5′-C/TNAG-3′
Ligation and recutting results: After 2-10-fold Dde I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >95% of fragments can be ligated, and >95% recut.
Heat inactivation: Completed after 65 °C incubation for 20 minutes.
Comment: Single-stranded DNA cleaves slowly.
10835307001 Dde I from Desulfovibrio desulfuricans, strain Norway Recognition sites: *CTNAG
*CTNAG
Restriction site: *C↓TNAG
*C↓TNAG
Heat inactivation: No inactivation of Dde I after incubation at 65 °C for 15 minutes.
R8381 Dpn I from Diplococcus pneumoniae Restriction Enzyme Recognition sequence: 5′-GmA/TC-3′
Ligation and recutting results: After 2-10-fold Dpn I overdigestion of 1 μg pBR322 DNA substrate, results in 100% cutting, >30% of fragments can be ligated, and >90% recut.
Heat inactivation: 75 °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.
R4381 Dra I from Deinococcus radiophilus Restriction Enzyme Recognition sequence: 5′-TTT/AAA-3′
Cutting results: a 2-10-fold Dra I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: 65 °C for 20 minutes.
10827754001 Dra I from Deinococcus radiophilus Recognition sites: TTTA°AA
TTTA°AA
Restriction site: TTT↓A°AA
TTT↓A°AA
Heat inactivation: No inactivation of Dra 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.
11167103001 Eco47 III from Escherichia coli RFL 47 Recognition sites: °AG*CGCT
°AG*CGCT
Restriction site: °AG*C↓GCT
°AG*C↓GCT
Heat inactivation: Eco47 III can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
R6265 EcoR I from Escherichia coli BS5 Restriction Enzyme Recognition sequence: 5′-G/AATTC-3′
Cutting results: a 2-10-fold EcoR I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: 65 °C for 20 minutes.
ECORI-RO EcoR I from Escherichia coli BS5 Star Activity
The enzyme shows star activity under non-optimal conditions. The recognition specificity of EcoR I is altered by addition of increasing amounts of hydrophobic reagents and glycerol to the incubation mixture. The relaxed specificities of EcoR I are N/AATTN or RRATYY. When digesting genomic DNA with high concentrated EcoR I note that a 1:10 dilution of enzyme solution is the minimum dilution.
Recognition sites: G*A*ATT*C
G*A*ATT*C
Restriction site: G↓*A*ATT*C
G↓*A*ATT*C
Heat inactivation: EcoR I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 10 U/μg DNA).
Star activity: To avoid star activity, always use the optimal buffer system and enzyme amount recommended in the package insert. Make sure that the DNA preparation is free of organic solvents and contaminating salts.
R2756 EcoR V from Escherichia coli buffered aqueous glycerol solution Recognition sequence: 5′-GAT/ATC-3′
Ligation and recutting results: After 2-10-fold Eco RV overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >85% of fragments can be ligated, and >95% recut.
Heat inactivation: 80 °C for 20 minutes.
ECORV-RO EcoR V from Escherichia coli J62 pLG74 Star Activity
EcoR V exhibits star activity under non-optimal conditions.
Recognition sites: G*ATAT °C
G*ATAT °C
Restriction site: G*AT↓AT °C
G*AT↓AT °C
Heat inactivation: No inactivation of EcoR V after incubation at 65 °C for 15 minutes.
R5628 Hae III from Haemophilus aegyptius Restriction Enzyme Recognition sequence: 5′-GG/CC-3′
Ligation and recutting results: After 2-10-fold Hae III overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >50% of fragments can be ligated, and >95% recut.
Heat inactivation: 80 °C for 20 minutes.
10693944001 Hae III from Haemophilus aegyptius Recognition sites: GG*C °C
GG*C °C
Restriction site: GG↓*C °C
GG↓*C °C
Heat inactivation: No inactivation of Hae III after incubation at 65 °C for 15 minutes.
R1137 Hind III from Haemophilus influenzae Restriction Enzyme Recognition sequence: 5′-A/AGCTT-3′
Cutting results: 2-10-fold Hind III overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: 65 °C for 15 minutes
HINDIII-RO Hind III from Haemophilus influenzae Rd com-10 Star Activity
Hind III exhibits star activity under non-optimal conditions. The recognition specificity of Hind III is altered by addition of increasing amounts of hydrophobic reagents and glycerol to the incubation mixture. This activity is the so-called star activity.
Recognition sites: AAGCTT
AAGCTT
Restriction site: *A↓°AG*CTT
*A↓°AG*CTT
Heat inactivation: Hind III can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
10656305001 Hind II from Haemophilus influenzae Rd com-10 Recognition sites: GTYRAC
GTYRACGT(T/C)(A/G)AC
Restriction site: GTY↓R*AC
GTY↓R*ACGT(T/C)↓(A/G)AC
Heat inactivation: Hind II can be heat inactivated by incubation at 65 °C for 15 minutes.
HINFI-RO Hinf I from Haemophilus influenzae Rf Recognition sites: GANTC
GANTC
Restriction site: G↓*ANT °C
G↓*ANT °C
Heat inactivation: No inactivation of Hinf I after incubation at 65 °C for 15 minutes.
R8507 Hpa I from Haemophilus parainfluenzae Restriction Enzyme Recognition sequence: 5′-GTT/AAC-3′
Ligation and recutting results: After 2-10-fold Hpa I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >80% of fragments can be ligated, and >95% recut.
Heat inactivation: Not completely inactivated at 65 °C for 15 minutes.
R0629 Hpa II from Haemophilus parainfluenzae buffered aqueous glycerol solution Recognition sequence: 5′-C/CGG-3′
Ligation and recutting results: After 2-10-fold Hpa II 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.
HPAI-RO Hpa I from Haemophilus parainfluenza Recognition sites: GTT*A°AC
GTT*A°AC
Restriction site: GTT↓*A°AC
GTT↓*A°AC
Heat inactivation: No inactivation of Hpa I after incubation at 65 °C for 15 minutes.
KPNI-RO Kpn I from Klebsiella pneumoniae OK8 The recognition specifiity of Kpn I is altered by addition of increasing amounts of hydrophobic reagents and glycerol to the incubation mixture.
Recognition sites: GGTACC
GGTACC
Restriction site: GGTAC↓C
GGTAC↓C
Heat inactivation: Kpn I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 3 U/μg DNA).
Star activity: Kpn I may exhibit star activity under non-optimal conditions.
11117807001 Ksp I (Sac II) from Kluyvera species Recognition sites: CCGCGG
CCGCGG
Restriction site: CCGC↓GG
CCGC↓GG
Heat inactivation: No inactivation of Ksp I after incubation at 65 °C for 15 minutes.
R4134 Ksp I from Kluyvera sp. Restriction Enzyme Recognition sequence: 5′-CCGC/GG-3′
Ligation and recutting results: After 2-10-fold Ksp I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >90% of fragments can be ligated, and >90% recut.
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.
MLUI-RO Mlu I from Micrococcus luteus Recognition sites: °A*CGCGT
°A*CGCGT
Restriction site: °A↓*CGCGT
°A↓*CGCGT
Heat inactivation: There is no information available whether or not Mlu I can be heat inactivated.
11102982001 Mro I (Acc III) from Micrococcus roseus Recognition sites: T↓*CCGG°A
T*CCGG°A
Restriction site: T↓*CCGG°A
T↓*CCGG°A
Heat inactivation: There is no information available whether or not Mvn I can be heat inactivated.
R4506 Msp I from Moraxella sp. buffered aqueous glycerol solution Recognition sequence: 5′-C/CGG-3′
Ligation and recutting results: After 2-10-fold Msp I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >95% of fragments can be ligated, and >95% recut.
Heat inactivation: Inactivated at 65 °C for 15 minutes.
11441337001 Mun I (Mfe I) from Mycoplasma species Star Activity
The recognition specificity of Mun I is relaxed in extended incubation with excess enzyme, or at low ionic strength.
Recognition sites: CA*ATTG
CA*ATTG
Restriction site: C↓A*ATTG
C↓A*ATTG
Heat inactivation: Mun I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 15 U/μg DNA).
R1632 Mva I from Micrococcus varians Rfl 19 Restriction Enzyme Recognition sequence: 5′-CC/(A,T)GG-3′
Ligation and recutting results: After 2-10-fold Mva I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >90% of fragments can be ligated, and >90% recut
Heat inactivation: Inactivated at 60 °C for 15 minutes.
11103024001 Nar I from Nocardia argentinensis Recognition sites: GG*CGCC
GG*CGCC
Restriction site: GG↓*CGCC
GG↓*CGCC
Heat inactivation: Nar I can be heat inactivated by incubation at 65 °C for 15 minutes.
R8761 Nco I from Nocardia corallina Restriction Enzyme Recognition sequence: 5′-C/CATGG-3′
Cutting results: a 2-10-fold Nco I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Inactivated at 65 °C for 15 minutes.
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).
R5509 Nde I from Neisseria denitrificans Restriction Enzyme Recognition sequence: 5′-CA/TATG-3′
Cutting results: a 2-10-fold Nde I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Inactivated at 60 °C for 15 minutes.
NDEI-RO Nde I fom Neisseria denitrificans NRCC 31009 Recognition sites: °CAT*ATG
°CAT*ATG
Restriction site: °CA↓T*ATG
°CA↓T*ATG
Heat inactivation: Nde I can be heat inactivated by incubation at 65 °C for 15 minutes.

Methylation sensitivity: Nde I is inhibited by internal 6-methyladenine at the site indicated (*) on the recognition sequence. The enzyme is not inhibited by 5-methylcytosine (°).
R5634 Nhe I from Neisseria mucosa heidelbergensis Restriction Enzyme Recognition sequence: 5′-G/CTAGC-3′
Cutting results: a 2-10-fold Nhe I overdigestion of 1 μg λ DNA substrate results in 100% cutting.
Heat inactivation: Inactivated at 65 °C for 20 minutes.
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.
R8506 Not I from Nocardia otidiscaviarum Restriction Enzyme Recognition sequence: 5′-GC/GGCCGC-3′
Cutting results: a 2-10-fold Not I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Inactivated at 65 °C for 15 minutes.
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).
10776777001 Nru I from Nocardia rubra Recognition sites: T*CG↓ °CG*A
T*CG↓ °CG*A
Restriction site: T*CG↓ °CG*A
T*CG↓ °CG*A
Heat inactivation: Nru I can be heat inactivated by incubation at 65 °C for 15 minutes.
R5884 Nsi I from Neisseria sicca Restriction Enzyme Recognition sequence: 5′-ATGCA/T-3′
Heat inactivation: Inactivated at 65 °C for 20 minutes.
NSII-RO Nsi I from Neisseria sicca Recognition sites: ATGC*AT
ATGC*AT
Restriction site: ATGC*A↓T
ATGC*A↓T
Heat inactivation: Nsi I can be heat inactivated by incubation at 65 °C for 15 minutes.
R7023 Pst I from Providencia stuartii Restriction Enzyme Recognition sequence: 5′-CTGCA/G-3′
Cutting results: a 2-10-fold Pst I overdigestion of 1 μg λ DNA substrate results in 100% cutting.
Heat inactivation: Inactivated at 80 °C for 20 minutes.
PSTI-RO Pst I from Providencia stuartii The recognition specificity of Pst I is altered by addition of increasing amounts of hydrophobic reagents and glycerol to the incubation mixture.
Recognition sites: CTGCAG
CTGCAG
Restriction site: CTGCA↓G
CTGCA↓G
Heat inactivation: No Inactivation of Pst I by incubation at 65 °C for 15 minutes.
R1508 Pvu I from Proteus vulgaris Restriction Enzyme Recognition sequence: 5′-CGAT/CG-3′
Cutting results: A 2-10-fold Pvu I overdigestion of 1 μg λ DNA substrate results in 100% cutting.
Heat inactivation: Inactivated at 80 °C for 20 minutes.
R2631 Pvu II from Proteus vulgaris Restriction Enzyme Recognition sequence: 5′-CAG/CTG-3′
Cutting results: a 2-10-fold Pvu II overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Activity not completely destroyed at 65 °C for 15 minutes.
PVUII-RO Pvu II from Proteus vulgaris Star Activity
Pvu II exhibits star activity under non-optimal conditions. The relaxed sequence specificity of Pvu II includes all sequence combinations which differ by one nucleotide from the non-relaxed sequence (e.g., CCGCTG, CATCTG, CAGATG).
Recognition sites: CAG*CTG
CAG*CTG
Restriction site: CAG↓*CTG
CAG↓*CTG
Heat inactivation: No inactivation of Pvu II after incubation at 65 °C for 15 minutes.
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.
R4756 Rsa I from Rhodopseudomonas sphaeroides Restriction Enzyme Recognition sequence: 5′-GT/AC-3′
Cutting results: a 2-10-fold Rsa I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Inactivated at 65 °C for 20 minutes.
RSAI-RO Rsa I from Rhodopseudomonas sphaeroides Recognition sites: GT*A*C
GT*A*C
Restriction site: GT↓*A*C
GT↓*A*C
Heat inactivation: Rsa I can be heat inactivated by incubation at 65 °C for 15 minutes.

Methylation sensitivity
Cleavage by Rsa I is not inhibited by the presence of 5-methylcytosine at the site (°) indicated on the recognition sequence. However, the enzyme is inhibited by 6-methyladenine (*) or 4-methylcytosine (°).

Activity in SuRE/Cut Buffer System
Buffer printed in bold face type is the buffer recommended for optimal activity:
A B< L M H
100% 50-75% 100% 50-75% 0-10%

Relative activity in complete PCR mix
Relative activity in PCR mix (Taq DNA Polymerase buffer) is 100%. The PCR mix contained λDNA, primers, 10 mM Tris-HCl (pH 8.3, +20°C), 50 mM KCl, 1.5 mM MgCl2, 200 μM dNTPs, 2.5 U Taq DNA polymerase. The mix was subjected to 25 amplification cycles.

Incubation temperature: +37°C

Unit definition
One unit is the enzyme activity that completely cleaves 1 μg ?DNA in 1 hour at +37°C in a total volume of 25 μl SuRE/Cut Buffer L. Complete digestion of 1 μg pBR322 DNA requires 1 U of Rsa I.

Heat inactivation
Rsa I can be heat inactivated by incubating it for 15 minutes at +65°C.

Number of cleavage sites on different DNAs
λ Ad2 SV40 FX174 M13mp7 M13mp18 pBR322 pBR328 pUC18
113 83 12 11 18 19 3 4 3

Ligation and recutting assay
Rsa I fragments obtained by complete digestion of 1 μg λDNA are ligated with 1 U T4 DNA Ligase in a volume of 10 μl by incubation for 16 hours at +4°C in 66 mM Tris-HCl, 5 mM MgCl2, 5 mM Dithiothreitol, 1 mM ATP, pH 7.5 (at +20°C) resulting in >50% recovery of 1 μg λDNA fragments.
Subsequent re-cutting with Rsa I yields >95% of the typical pattern of λDNA × Rsa I fragments.

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).
R5268 Sac I from Streptomyces achromogenes Restriction Enzyme Recognition sequence: 5′-GAGCT/C-3′
Cutting results: a 2-10-fold Sac I overdigestion of 1 μg λ Hind III DNA substrate results in 100% cutting
Heat inactivation : Inactivated at 65 °C for 15 minutes.
R0754 Sal I from Streptomyces albus G Restriction Enzyme Recognition sequence: 5′-G/TCGAC-3′
Cutting results: a 2-10-fold Sal I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Inactivated at 65 °C for 15 minutes.
SALI-RO Sal I from Streptomyces albus G Star Activity
The most common star activity is the relaxed specificity, i.e. the enzyme not only recognizes the 6 bp palindrome but also the 4 bp core sequence. Sal I exhibits star activity in low salt (buffer H is recommended) and in high glycerol. pH also appears to play a role in exhibition of star activity: pH 8.0 is already too high. Do not leave Sal I at room temperature (15 to 25 °C), not even for a short period.
Recognition sites: GT*CG*AC
GT*CG*AC
Restriction site: G↓T*CG*AC
G↓T*CG*AC
Heat inactivation: Sal I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
R0762 Sau3A I from Staphylococcus aureus Restriction Enzyme Recognition sequence: 5′-/GATC-3′
Cutting results: A 2-10-fold Sau3A I overdigestion of 1 μg λ DNA substrate results in 100% cutting.
Heat inactivation: 65 °C for 20 minutes.
R5007 Sca I from Streptomyces caespitosus Restriction Enzyme Recognition sequence: 5′-AGT/ACT-3′
Ligation and recutting results: After 2-10-fold Sca I overdigestion of 1 μg λ DNA substrate, results in 100% cutting, >90% of fragments can be ligated, and >90% recut.
Heat inactivation: Inactivated at 80 °C for 20 minutes.
10775266001 Sca I from Streptoverticillium species Star Activity
The recognition specificity of Sca I is relaxed at low ionic strength or by increasing amounts of enzyme or by addition of glycerol to the incubation mixture.
Recognition sites: AGTA °CT
AGTA °CT
Restriction site: AGT↓A °CT
AGT↓A °CT
Heat inactivation: No inactivation of Sca I after incubation at 65 °C for 15 minutes.
R8256 Sfi I from Streptomyces fimbriatus Restriction Enzyme Recognition sequence: 5′-GGCC(N)4/NGGCC-3′
Cutting results: a 2-10-fold Sfi I overdigestion of 1 μg Ad-2 DNA substrate results in 100% cutting
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
11243497001 Sfu I (Asu II) from Streptomyces fulvissimus Recognition sites: TT °CG*AA
TT °CG*AA
Restriction site: TT↓ °CG*AA
TT↓ °CG*AA
Heat inactivation: There is no information available whether or not Sfu I can be heat inactivated.
R4503 Sma I from Serratia marcescens Sb Restriction Enzyme Recognition sequence: 5′-CCC/GGG-3′
Cutting results: a 2-10-fold Sma I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: 65 °C for 15 minutes.
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).
R5257 Spe I from Sphaerotilus sp. Restriction Enzyme Recognition sequence: 5′-A/CTAGT-3′
Cutting results: a 2-10-fold Spe I overdigestion of 1 μg λ DNA substrate results in 100% cutting.
Heat inactivation: Inactivated at 65 °C for 20 minutes.
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).
R7135 Sph I from Streptomyces phaeochromogenes Restriction Enzyme Recognition sequence: 5′-GCATG/C-3′
Cutting results: A 2-10-fold Sph I overdigestion of 1 μg λ DNA substrate results in 100% cutting.
Heat inactivation: Inactivated at 65 °C for 20 minutes.
SPHI-RO Sph I from Streptomyces phaeochromogenes Recognition sites: GC*ATG °C
GC*ATG °C
Restriction site: GC*ATG↓ °C
GC*ATG↓ °C
Heat inactivation: Sph I can be heat inactivated by incubation at 65 °C for 15 minutes (up to 100 U/μg DNA).
STUI-RO Stu I from Streptomyces nodosus Recognition sites: AGG*C*CT
AGG*C*CT
Restriction site: AGG↓*C*CT
AGG↓*C*CT
Heat inactivation: Stu I can be heat inactivated by incubation at 65 °C for 15 minutes.
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.
R9507 Taq I from Thermus aquaticus recombinant, expressed in E. coli (Strain that carries a Taq I overproducing plasmid.), Restriction Enzyme Recognition sequence: 5′-T/CGA-3′
Ligation and recutting results: After 2-10-fold Taq I overdigestion of 1 μg λ DNA substrate, results in >95% cutting, >85% of fragments can be ligated, and >95% recut.
Heat inactivation: 80 °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.
R7260 Xba I from Xanthomonas badrii Restriction Enzyme Recognition sequence: 5′-T/CTAGA-3′
Cutting results: A 2-10-fold Xba I overdigestion of 1 μg Ad-2 DNA substrate results in 100% cutting
Heat inactivation: Up to 15 units of enzyme inactivated 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.
R6379 Xho I from Xanthomonas holcicola Restriction Enzyme Recognition sequence: 5′-C/TCGAG-3′
Cutting results: a 2-10-fold Xho I overdigestion of 1 μg λ DNA substrate results in 100% cutting
Heat inactivation: Inactivated at 65 °C for 15 minutes.
XHOI-RO Xho I from Xanthomonas campestris (formerly Xanthomonas holcicola) Recognition sites: *CT*CG*AG
*CT*CG*AG
Restriction site: *C↓T*CG*AG
*C↓T*CG*AG
Heat inactivation: No inactivation of Xho I after incubation at 65 °C for 15 minutes. It is recommended to inactivate Xho I by phenolization.
Relative activity in complete PCR mix
Relative activity in PCR mix (Taq DNA Polymerase buffer) is <5%. The PCR mix contained ?DNA, primers, 10 mM Tris-HCl (pH 8.3, +20°C), 50 mM KCl, 1.5 mM MgCl2, 200 μM dNTPs, 2.5 U Taq DNA polymerase. The mix was subjected to 25 amplification cycles. Activity in reaction buffer of Pwo SuperYield DNA Polymerase PCR Mix is 15%. When supplemented with GC-RICH Solution, activity remains at 15%.
Incubation temperature
+37°C