NEB’s restriction enzyme buffer system makes your restriction digests easy and convenient. We are able to offer >210 restriction enzymes that cut in a single buffer, CutSmart® .
This improves ease-of-use, especially when performing double digests. In addition to indicating the performance of each enzyme in the 4 NEBuffers, the chart also
indicates ligation and recutting, star activity, and whether or not more than 1-site is required for cleavage.
Should you require information on the pre-2013 buffer system, please refer to the old NEBuffer Activity Chart. You can also receive additional support by contacting info@neb.com.
Enzyme | Sequence | Supplied NEBuffer | % Activity in NEBuffer | Heat Inac. | Incu. Temp. | Diluent | |||
1.1 | 2.1 | 3.1 | CutSmart | ||||||
AatII | GACGT/C | CutSmart® Buffer | 10 | 50* | 50 | 100 | 80°C | 37°C | B |
AbaSI | CNNNNNNNNNNN/NNNNNNNNNG | CutSmart® Buffer | 25 | 50 | 50 | 100 | 65°C | 25°C | C |
Acc65I | G/GTACC | NEBuffer™ 3.1 | 10 | 75* | 100 | 25 | 65°C | 37°C | A |
AccI | GT/MKAC | CutSmart® Buffer | 50 | 50 | 10 | 100 | 80°C | 37°C | A |
AciI | CCGC(-3/-1) | CutSmart® Buffer | 10 | 25 | 100 | 100 | 65°C | 37°C | A |
AclI | AA/CGTT | CutSmart® Buffer | 10 | 10 | 10 | 100 | No | 37°C | B |
AcuI | CTGAAG(16/14) | CutSmart® Buffer + SAM | 50 | 100 | 50 | 100 | 65°C | 37°C | B |
AfeI | AGC/GCT | CutSmart® Buffer | 25 | 100 | 25 | 100 | 65°C | 37°C | B |
AflII | C/TTAAG | CutSmart® Buffer | 50 | 100 | 10 | 100 | 65°C | 37°C | A |
AflIII | A/CRYGT | NEBuffer™ 3.1 | 10 | 50 | 100 | 50 | 80°C | 37°C | B |
AgeI § | A/CCGGT | NEBuffer™ 1.1 | 100 | 75 | 25 | 75 | 65°C | 37°C | C |
AgeI-HF® | A/CCGGT | CutSmart® Buffer | 100 | 50 | 10 | 100 | 65°C | 37°C | A |
AhdI | GACNNN/NNGTC | CutSmart® Buffer | 25 | 25 | 10 | 100 | 65°C | 37°C | A |
AleI-v2 | CACNN/NNGTG | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
AluI | AG/CT | CutSmart® Buffer | 25 | 100 | 50 | 100 | 80°C | 37°C | B |
AlwI | GGATC(4/5) | CutSmart® Buffer | 50 | 50 | 10 | 100 | No | 37°C | A |
AlwNI | CAGNNN/CTG | CutSmart® Buffer | 10 | 100 | 50 | 100 | 80°C | 37°C | A |
ApaI | GGGCC/C | CutSmart® Buffer | 25 | 25 | 10 | 100 | 65°C | 25°C | A |
ApaLI | G/TGCAC | CutSmart® Buffer | 100 | 100 | 10 | 100 | No | 37°C | A |
ApeKI | G/CWGC | NEBuffer™ 3.1 | 25 | 50 | 100 | 10 | No | 75°C | B |
ApoI § | R/AATTY | NEBuffer™ 3.1 | 10 | 75 | 100 | 75 | 80°C | 50°C | A |
ApoI-HF | R/AATTY | CutSmart® Buffer | 10 | 100 | 10 | 100 | 80°C | 37°C | B |
AscI | GG/CGCGCC | CutSmart® Buffer | 10 | 10 | 10 | 100 | 80°C | 37°C | A |
AseI | AT/TAAT | NEBuffer™ 3.1 | 10 | 50* | 100 | 10 | 65°C | 37°C | B |
AsiSI | GCGAT/CGC | CutSmart® Buffer | 100 | 100 | 25 | 100 | 80°C | 37°C | B |
AvaI | C/YCGRG | CutSmart® Buffer | 10 | 100 | 25 | 100 | 80°C | 37°C | A |
AvaII | G/GWCC | CutSmart® Buffer | 50 | 75 | 10 | 100 | 80°C | 37°C | A |
AvrII | C/CTAGG | CutSmart® Buffer | 100 | 50 | 50 | 100 | No | 37°C | B |
BaeGI | GKGCM/C | NEBuffer™ 3.1 | 75 | 75 | 100 | 25 | 80°C | 37°C | A |
BaeI | (10/15)ACNNNNGTAYC(12/7) | CutSmart® Buffer + SAM | 50 | 100 | 50 | 100 | 65°C | 25°C | A |
BamHI § | G/GATCC | NEBuffer™ 3.1 | 75* | 100* | 100 | 100* | No | 37°C | A |
BamHI-HF® | G/GATCC | CutSmart® Buffer | 100 | 50 | 10 | 100 | No | 37°C | A |
BanI | G/GYRCC | CutSmart® Buffer | 10 | 25 | 10 | 100 | 65°C | 37°C | A |
BanII | GRGCY/C | CutSmart® Buffer | 100 | 100 | 50 | 100 | 80°C | 37°C | A |
BbsI § | GAAGAC(2/6) | NEBuffer™ 2.1 | 100 | 100 | 25 | 75 | 65°C | 37°C | B |
BbsI-HF® | GAAGAC(2/6) | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
BbvCI | CCTCAGC(-5/-2) | CutSmart® Buffer | 10 | 100 | 50 | 100 | No | 37°C | B |
BbvI | GCAGC(8/12) | CutSmart® Buffer | 100 | 100 | 25 | 100 | 65°C | 37°C | B |
BccI | CCATC(4/5) | CutSmart® Buffer | 100 | 50 | 10 | 100 | 65°C | 37°C | A |
BceAI | ACGGC(12/14) | NEBuffer™ 3.1 | 100* | 100* | 100 | 100* | 65°C | 37°C | A |
BcgI | (10/12)CGANNNNNNTGC(12/10) | NEBuffer™ 3.1 + SAM | 10 | 75* | 100 | 50* | 65°C | 37°C | A |
BciVI | GTATCC(6/5) | CutSmart® Buffer | 100 | 25 | 10 | 100 | 80°C | 37°C | C |
BclI § | T/GATCA | NEBuffer™ 3.1 | 50 | 100 | 100 | 75 | No | 50°C | A |
BclI-HF | T/GATCA | CutSmart® Buffer | 100 | 100 | 10 | 100 | 65°C | 37°C | B |
BcoDI | GTCTC(1/5) | CutSmart® Buffer | 50 | 75 | 75 | 100 | No | 37°C | B |
BfaI | C/TAG | CutSmart® Buffer | 10 | 10 | 10 | 100 | 80°C | 37°C | B |
BfuAI | ACCTGC(4/8) | NEBuffer™ 3.1 | 10 | 25 | 100 | 10 | 65°C | 50°C | B |
BglI | GCCNNNN/NGGC | NEBuffer™ 3.1 | 10 | 25 | 100 | 10 | 65°C | 37°C | B |
BglII | A/GATCT | NEBuffer™ 3.1 | 10 | 10 | 100 | 10 | No | 37°C | A |
BlpI | GC/TNAGC | CutSmart® Buffer | 50 | 100 | 10 | 100 | No | 37°C | A |
BmgBI | CACGTC(-3/-3) | NEBuffer™ 3.1 | 10 | 10 | 100 | 10 | 65°C | 37°C | B |
BmrI | ACTGGG(5/4) | NEBuffer™ 2.1 | 75 | 100 | 75 | 100* | 65°C | 37°C | B |
BmtI § | GCTAG/C | NEBuffer™ 3.1 | 100 | 100 | 100 | 100 | 65°C | 37°C | B |
BmtI-HF® | GCTAG/C | CutSmart® Buffer | 50 | 100 | 10 | 100 | 65°C | 37°C | B |
BpmI | CTGGAG(16/14) | NEBuffer™ 3.1 | 75 | 100 | 100 | 100 | 65°C | 37°C | B |
BpuEI | CTTGAG(16/14) | CutSmart® Buffer + SAM | 50* | 100 | 50* | 100 | 65°C | 37°C | B |
Bpu10I | CCTNAGC(-5/-2) | NEBuffer™ 3.1 | 10 | 25 | 100 | 25 | 80°C | 37°C | B |
BsaAI | YAC/GTR | CutSmart® Buffer | 100 | 100 | 100 | 100 | No | 37°C | C |
BsaBI | GATNN/NNATC | CutSmart® Buffer | 50 | 100 | 75 | 100 | 80°C | 60°C | B |
BsaHI | GR/CGYC | CutSmart® Buffer | 50 | 100 | 100 | 100 | 80°C | 37°C | C |
BsaI § | GGTCTC(1/5) | CutSmart® Buffer | 75* | 75 | 100 | 100 | 65°C | 37°C | B |
BsaI-HF®v2 | GGTCTC(1/5) | CutSmart® Buffer | 100 | 100 | 100 | 100 | 80°C | 37°C | B |
BsaJI | C/CNNGG | CutSmart® Buffer | 50 | 100 | 100 | 100 | 80°C | 60°C | A |
BsaWI | W/CCGGW | CutSmart® Buffer | 10 | 100 | 50 | 100 | 80°C | 60°C | A |
BsaXI | (9/12)ACNNNNNCTCC(10/7) | CutSmart® Buffer | 50* | 100* | 10 | 100 | No | 37°C | C |
BseRI | GAGGAG(10/8) | CutSmart® Buffer | 100 | 100 | 75 | 100 | 80°C | 37°C | A |
BseYI | CCCAGC(-5/-1) | NEBuffer™ 3.1 | 10 | 50 | 100 | 50 | 80°C | 37°C | B |
BsgI | GTGCAG(16/14) | CutSmart® Buffer + SAM | 25 | 50 | 25 | 100 | 65°C | 37°C | B |
BsiEI | CGRY/CG | CutSmart® Buffer | 25 | 50 | 10 | 100 | No | 60°C | A |
BsiHKAI | GWGCW/C | CutSmart® Buffer | 25 | 100 | 100 | 100 | No | 65°C | A |
BsiWI § | C/GTACG | NEBuffer™ 3.1 | 25 | 50* | 100 | 25 | 65°C | 55°C | B |
BsiWI-HF® | C/GTACG | CutSmart® Buffer | 50 | 100 | 10 | 100 | No | 37°C | B |
BslI | CCNNNNN/NNGG | CutSmart® Buffer | 50 | 75 | 100 | 100 | No | 55°C | A |
BsmAI | GTCTC(1/5) | CutSmart® Buffer | 50 | 100 | 100 | 100 | No | 55°C | B |
BsmBI | CGTCTC(1/5) | NEBuffer™ 3.1 | 10 | 50* | 100 | 25 | 80°C | 55°C | B |
BsmFI | GGGAC(10/14) | CutSmart® Buffer | 25 | 50 | 50 | 100 | 80°C | 65°C | A |
BsmI | GAATGC(1/-1) | CutSmart® Buffer | 25 | 100 | 10 | 100 | 80°C | 65°C | A |
BsoBI | C/YCGRG | CutSmart® Buffer | 25 | 100 | 100 | 100 | 80°C | 37°C | A |
BspCNI | CTCAG(9/7) | CutSmart® Buffer + SAM | 100 | 75 | 10 | 100 | 80°C | 25°C | A |
BspDI | AT/CGAT | CutSmart® Buffer | 25 | 75 | 50 | 100 | 80°C | 37°C | A |
BspEI | T/CCGGA | NEBuffer™ 3.1 | 10 | 10 | 100 | 10 | 80°C | 37°C | B |
BspHI | T/CATGA | CutSmart® Buffer | 10 | 50 | 25 | 100 | 80°C | 37°C | A |
Bsp1286I | GDGCH/C | CutSmart® Buffer | 25 | 25 | 25 | 100 | 65°C | 37°C | A |
BspMI | ACCTGC(4/8) | NEBuffer™ 3.1 | 10 | 50* | 100 | 10 | 65°C | 37°C | B |
BspQI | GCTCTTC(1/4) | NEBuffer™ 3.1 | 100* | 100* | 100 | 100* | 80°C | 50°C | B |
BsrBI | CCGCTC(-3/-3) | CutSmart® Buffer | 50 | 100 | 100 | 100 | 80°C | 37°C | A |
BsrDI | GCAATG(2/0) | NEBuffer™ 2.1 | 10 | 100 | 75 | 25 | 80°C | 65°C | A |
BsrFI-v2 | R/CCGGY | CutSmart® Buffer | 25 | 25 | 0 | 100 | No | 37°C | C |
BsrGI § | T/GTACA | NEBuffer™ 2.1 | 25 | 100 | 100 | 25 | 80°C | 37°C | A |
BsrGI-HF® | T/GTACA | CutSmart® Buffer | 10 | 100 | 100 | 100 | 80°C | 37°C | A |
BsrI | ACTGG(1/-1) | NEBuffer™ 3.1 | 10 | 50 | 100 | 10 | 80°C | 65°C | B |
BssHII | G/CGCGC | CutSmart® Buffer | 100 | 100 | 100 | 100 | 65°C | 50°C | B |
BssSI-v2 | CACGAG(-5/-1) | CutSmart® Buffer | 10 | 25 | 10 | 100 | No | 37°C | B |
BstAPI | GCANNNN/NTGC | CutSmart® Buffer | 50 | 100 | 25 | 100 | 80°C | 60°C | A |
BstBI | TT/CGAA | CutSmart® Buffer | 75 | 100 | 10 | 100 | No | 65°C | A |
BstEII § | G/GTNACC | NEBuffer™ 3.1 | 10 | 75* | 100 | 75* | No | 60°C | A |
BstEII-HF® | G/GTNACC | CutSmart® Buffer | 10 | 10 | 10 | 100 | No | 37°C | A |
BstNI | CC/WGG | NEBuffer™ 3.1 | 10 | 100 | 100 | 75 | No | 60°C | A |
BstUI | CG/CG | CutSmart® Buffer | 50 | 100 | 25 | 100 | No | 60°C | A |
BstXI | CCANNNNN/NTGG | NEBuffer™ 3.1 | 10 | 50 | 100 | 25 | 80°C | 37°C | B |
BstYI | R/GATCY | NEBuffer™ 2.1 | 25 | 100 | 75 | 100 | No | 60°C | A |
BstZ17I-HF® | GTATAC | CutSmart® Buffer | 100 | 100 | 10 | 100 | No | 37°C | A |
Bsu36I | CC/TNAGG | CutSmart® Buffer | 25 | 100 | 100 | 100 | 80°C | 37°C | C |
BtgI | C/CRYGG | CutSmart® Buffer | 50 | 100 | 100 | 100 | 80°C | 37°C | B |
BtgZI | GCGATG(10/14) | CutSmart® Buffer | 10 | 25 | 10 | 100 | 80°C | 60°C | A |
BtsCI | GGATG(2/0) | CutSmart® Buffer | 10 | 100 | 25 | 100 | 80°C | 50°C | B |
BtsIMutI | CAGTG(2/0) | CutSmart® Buffer | 100 | 50 | 10 | 100 | 80°C | 55°C | A |
BtsI-v2 | GCAGTG(2/0) | CutSmart® Buffer | 100 | 100 | 25 | 100 | No | 55°C | A |
Cac8I | GCN/NGC | CutSmart® Buffer | 50 | 75 | 100 | 100 | 65°C | 37°C | B |
ClaI | AT/CGAT | CutSmart® Buffer | 10 | 50 | 50 | 100 | 65°C | 37°C | A |
CspCI | (11/13)CAANNNNNGTGG(12/10) | CutSmart® Buffer + SAM | 10 | 100 | 10 | 100 | 65°C | 37°C | A |
CviAII | C/ATG | CutSmart® Buffer | 50 | 50 | 10 | 100 | 65°C | 25°C | C |
CviKI-1 | RG/CY | CutSmart® Buffer | 25 | 100 | 100 | 100 | No | 37°C | A |
CviQI | G/TAC | NEBuffer™ 3.1 | 75 | 100* | 100 | 75* | No | 25°C | C |
DdeI | C/TNAG | CutSmart® Buffer | 75 | 100 | 100 | 100 | 65°C | 37°C | B |
DpnI | GA/TC | CutSmart® Buffer | 100 | 100 | 75 | 100 | 80°C | 37°C | B |
DpnII | /GATC | NEBuffer™ DpnII | 25 | 25 | 100* | 25 | 65°C | 37°C | B |
DraI | TTT/AAA | CutSmart® Buffer | 75 | 75 | 50 | 100 | 65°C | 37°C | A |
DraIII-HF® | CACNNN/GTG | CutSmart® Buffer | 10 | 50 | 10 | 100 | No | 37°C | B |
DrdI | GACNNNN/NNGTC | CutSmart® Buffer | 25 | 50 | 10 | 100 | 65°C | 37°C | A |
EaeI | Y/GGCCR | CutSmart® Buffer | 10 | 50 | 10 | 100 | 65°C | 37°C | A |
EagI § | C/GGCCG | NEBuffer™ 3.1 | 10 | 25 | 100 | 10 | 65°C | 37°C | B |
EagI-HF® | C/GGCCG | CutSmart® Buffer | 25 | 100 | 100 | 100 | 65°C | 37°C | B |
EarI | CTCTTC(1/4) | CutSmart® Buffer | 50 | 10 | 10 | 100 | 65°C | 37°C | B |
EciI | GGCGGA(11/9) | CutSmart® Buffer | 100 | 50 | 50 | 100 | 65°C | 37°C | A |
Eco53kI | GAG/CTC | CutSmart® Buffer | 100 | 100 | 10 | 100 | 65°C | 37°C | A |
EcoNI | CCTNN/NNNAGG | CutSmart® Buffer | 50 | 100 | 75 | 100 | 65°C | 37°C | A |
EcoO109I | RG/GNCCY | CutSmart® Buffer | 50 | 100 | 50 | 100 | 65°C | 37°C | A |
EcoP15I | CAGCAG(25/27) | NEBuffer™ 3.1 + ATP | 75 | 100 | 100 | 100 | 65°C | 37°C | A |
EcoRI § | G/AATTC | NEBuffer™ EcoRI | 25 | 100* | 50 | 50* | 65°C | 37°C | C |
EcoRI-HF® | G/AATTC | CutSmart® Buffer | 10 | 100 | 10 | 100 | 65°C | 37°C | C |
EcoRV § | GAT/ATC | NEBuffer™ 3.1 | 10 | 50 | 100 | 10 | 80°C | 37°C | A |
EcoRV-HF® | GAT/ATC | CutSmart® Buffer | 25 | 100 | 100 | 100 | 65°C | 37°C | B |
Esp3I | CGTCTC(1/5) | CutSmart® Buffer | 100 | 100 | 10 | 100 | 65°C | 37°C | B |
FatI | 0 | NEBuffer™ 2.1 | 10 | 100 | 50 | 50 | 80°C | 55°C | A |
FauI | CCCGC(4/6) | CutSmart® Buffer | 100 | 50 | 10 | 100 | 65°C | 55°C | A |
Fnu4HI | GC/NGC | CutSmart® Buffer | 10 | 10 | 10 | 100 | No | 37°C | A |
FokI | GGATG(9/13) | CutSmart® Buffer | 100 | 100 | 75 | 100 | 65°C | 37°C | A |
FseI | GGCCGG/CC | CutSmart® Buffer | 100 | 75 | 10 | 100 | 65°C | 37°C | B |
FspEI | CC(12/16) | CutSmart® Buffer | 10 | 10 | 10 | 100 | 80°C | 37°C | B |
FspI | TGC/GCA | CutSmart® Buffer | 10 | 100 | 10 | 100 | No | 37°C | C |
HaeII | RGCGC/Y | CutSmart® Buffer | 25 | 100 | 10 | 100 | 80°C | 37°C | A |
HaeIII | GG/CC | CutSmart® Buffer | 50 | 100 | 25 | 100 | 80°C | 37°C | A |
HgaI | GACGC(5/10) | NEBuffer™ 1.1 | 100 | 100 | 25 | 100 | 65°C | 37°C | A |
HhaI | GCG/C | CutSmart® Buffer | 25 | 100 | 100 | 100 | 65°C | 37°C | A |
HincII | GTY/RAC | NEBuffer™ 3.1 | 25 | 100 | 100 | 100 | 65°C | 37°C | B |
HindIII § | A/AGCTT | NEBuffer™ 2.1 | 25 | 100 | 50 | 50 | 80°C | 37°C | B |
HindIII-HF® | A/AGCTT | CutSmart® Buffer | 10 | 100 | 10 | 100 | 80°C | 37°C | B |
HinfI | G/ANTC | CutSmart® Buffer | 50 | 100 | 100 | 100 | 80°C | 37°C | A |
HinP1I | G/CGC | CutSmart® Buffer | 100 | 100 | 100 | 100 | 65°C | 37°C | A |
HpaI | GTT/AAC | CutSmart® Buffer | 10 | 75* | 25 | 100 | No | 37°C | A |
HpaII | C/CGG | CutSmart® Buffer | 100 | 50 | 10 | 100 | 80°C | 37°C | A |
HphI | GGTGA(8/7) | CutSmart® Buffer | 50 | 50 | 10 | 100 | 65°C | 37°C | B |
HpyAV | CCTTC(6/5) | CutSmart® Buffer | 100 | 100 | 25 | 100 | 65°C | 37°C | 0 |
HpyCH4III | ACN/GT | CutSmart® Buffer | 100 | 25 | 10 | 100 | 65°C | 37°C | A |
HpyCH4IV | A/CGT | CutSmart® Buffer | 100 | 50 | 25 | 100 | 65°C | 37°C | A |
HpyCH4V | TG/CA | CutSmart® Buffer | 50 | 50 | 25 | 100 | 65°C | 37°C | A |
Hpy188I | TCN/GA | CutSmart® Buffer | 25 | 100 | 50 | 100 | 65°C | 37°C | A |
Hpy99I | CGWCG/ | CutSmart® Buffer | 50 | 10 | 10 | 100 | 65°C | 37°C | A |
Hpy166II | GTN/NAC | CutSmart® Buffer | 100 | 100 | 50 | 100 | 65°C | 37°C | C |
Hpy188III | TC/NNGA | CutSmart® Buffer | 100 | 100 | 10 | 100 | 65°C | 37°C | B |
I-CeuI | TAACTATAACGGTCCTAAGGTAGCGAA(-9/-13) | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
I-SceI | TAGGGATAACAGGGTAAT(-9/-13) | CutSmart® Buffer | 10 | 50 | 25 | 100 | 65°C | 37°C | B |
KasI | G/GCGCC | CutSmart® Buffer | 50 | 100 | 50 | 100 | 65°C | 37°C | B |
KpnI § | GGTAC/C | NEBuffer™ 1.1 | 100 | 75 | 10 | 50* | No | 37°C | A |
KpnI-HF® | GGTAC/C | CutSmart® Buffer | 100 | 25 | 10 | 100 | No | 37°C | A |
LpnPI | CCDG(10/14) | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
MboI | /GATC | CutSmart® Buffer | 75 | 100 | 100 | 100 | 65°C | 37°C | A |
MboII | GAAGA(8/7) | CutSmart® Buffer | 100* | 100 | 50 | 100 | 65°C | 37°C | C |
MfeI § | C/AATTG | CutSmart® Buffer | 75 | 50 | 10 | 100 | No | 37°C | A |
MfeI-HF® | C/AATTG | CutSmart® Buffer | 75 | 25 | 10 | 100 | No | 37°C | A |
MluCI | /AATT | CutSmart® Buffer | 100 | 10 | 10 | 100 | No | 37°C | A |
MluI § | A/CGCGT | NEBuffer™ 3.1 | 10 | 50 | 100 | 25 | 80°C | 37°C | A |
MluI-HF® | A/CGCGT | CutSmart® Buffer | 25 | 100 | 100 | 100 | No | 37°C | A |
MlyI | GAGTC(5/5) | CutSmart® Buffer | 50 | 50 | 10 | 100 | 65°C | 37°C | A |
MmeI | TCCRAC(20/18) | CutSmart® Buffer + SAM | 50 | 100 | 50 | 100 | 65°C | 37°C | B |
MnlI | CCTC(7/6) | CutSmart® Buffer | 75 | 100 | 50 | 100 | 65°C | 37°C | B |
MscI | TGG/CCA | CutSmart® Buffer | 25 | 100 | 100 | 100 | 80°C | 37°C | C |
MseI | T/TAA | CutSmart® Buffer | 75 | 100 | 75 | 100 | 65°C | 37°C | A |
MslI | CAYNN/NNRTG | CutSmart® Buffer | 50 | 50 | 10 | 100 | 80°C | 37°C | A |
MspA1I | CMG/CKG | CutSmart® Buffer | 10 | 50 | 10 | 100 | 65°C | 37°C | B |
MspI | C/CGG | CutSmart® Buffer | 75 | 100 | 50 | 100 | No | 37°C | A |
MspJI | CNNR(9/13) | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
MwoI | GCNNNNN/NNGC | CutSmart® Buffer | 10 | 100 | 100 | 100 | No | 60°C | B |
NaeI | GCC/GGC | CutSmart® Buffer | 25 | 25 | 10 | 100 | No | 37°C | A |
NarI | GG/CGCC | CutSmart® Buffer | 100 | 100 | 10 | 100 | 65°C | 37°C | A |
Nb.BbvCI | 0 | CutSmart® Buffer | 25 | 100 | 100 | 100 | 80°C | 37°C | A |
Nb.BsmI | 0 | NEBuffer™ 3.1 | 10 | 50 | 100 | 10 | 80°C | 65°C | A |
Nb.BsrDI | 0 | CutSmart® Buffer | 25 | 100 | 100 | 100 | 80°C | 65°C | A |
Nb.BssSI | 0 | NEBuffer™ 3.1 | 10 | 100 | 100 | 25 | No | 37°C | B |
Nb.BtsI | 0 | CutSmart® Buffer | 75 | 100 | 75 | 100 | 80°C | 37°C | A |
NciI | CC/SGG | CutSmart® Buffer | 100 | 25 | 10 | 100 | No | 37°C | A |
NcoI § | C/CATGG | NEBuffer™ 3.1 | 100 | 100 | 100 | 100 | 80°C | 37°C | A |
NcoI-HF® | C/CATGG | CutSmart® Buffer | 50 | 100 | 10 | 100 | 80°C | 37°C | B |
NdeI | CA/TATG | CutSmart® Buffer | 75 | 100 | 100 | 100 | 65°C | 37°C | A |
NgoMIV | G/CCGGC | CutSmart® Buffer | 100 | 50 | 10 | 100 | No | 37°C | A |
NheI § | G/CTAGC | NEBuffer™ 2.1 | 100 | 100 | 10 | 100 | 65°C | 37°C | C |
NheI-HF® | G/CTAGC | CutSmart® Buffer | 100 | 25 | 10 | 100 | 80°C | 37°C | C |
NlaIII | CATG/ | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
NlaIV | GGN/NCC | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
NmeAIII | GCCGAG(21/19) | CutSmart® Buffer + SAM | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
NotI § | GC/GGCCGC | NEBuffer™ 3.1 | 10 | 50 | 100 | 25 | 65°C | 37°C | C |
NotI-HF® | GC/GGCCGC | CutSmart® Buffer | 25 | 100 | 25 | 100 | 65°C | 37°C | A |
NruI § | TCG/CGA | NEBuffer™ 3.1 | 10 | 10 | 100 | 10 | No | 37°C | A |
NruI-HF® | TCG/CGA | CutSmart® Buffer | 0 | 25 | 50 | 100 | No | 37°C | A |
NsiI § | ATGCA/T | NEBuffer™ 3.1 | 10 | 75 | 100 | 25 | 65°C | 37°C | B |
NsiI-HF® | ATGCA/T | CutSmart® Buffer | 10 | 20 | 10 | 100 | 80°C | 37°C | B |
NspI | RCATG/Y | CutSmart® Buffer | 100 | 100 | 10 | 100 | 65°C | 37°C | A |
Nt.AlwI | GGATC(4/-5) | CutSmart® Buffer | 10 | 100 | 100 | 100 | 80°C | 37°C | A |
Nt.BbvCI | CCTCAGC(-5/-7) | CutSmart® Buffer | 50 | 100 | 10 | 100 | 80°C | 37°C | A |
Nt.BsmAI | GTCTC(1/-5) | CutSmart® Buffer | 100 | 50 | 10 | 100 | 65°C | 37°C | A |
Nt.BspQI | GCTCTTC(1/-7) | NEBuffer™ 3.1 | 10 | 25 | 100 | 10 | 80°C | 50°C | B |
Nt.BstNBI | GAGTC(4/-5) | NEBuffer™ 3.1 | 0 | 10 | 100 | 10 | 80°C | 55°C | A |
Nt.CviPII | (0/-1)CCD | CutSmart® Buffer | 10 | 100 | 25 | 100 | 65°C | 37°C | A |
PacI | TTAAT/TAA | CutSmart® Buffer | 100 | 75 | 10 | 100 | 65°C | 37°C | A |
PaeR7I | C/TCGAG | CutSmart® Buffer | 25 | 100 | 10 | 100 | No | 37°C | A |
PciI | A/CATGT | NEBuffer™ 3.1 | 50 | 75 | 100 | 50* | 80°C | 37°C | B |
PflFI | GACN/NNGTC | CutSmart® Buffer | 25 | 100 | 25 | 100 | 65°C | 37°C | A |
PflMI | CCANNNN/NTGG | NEBuffer™ 3.1 | 0 | 100 | 100 | 50 | 65°C | 37°C | A |
PI-PspI | TGGCAAACAGCTATTATGGGTATTATGGGT(-13/-17) | NEBuffer™ PI-PspI + BSA | 10 | 10 | 10 | 10 | No | 65°C | B |
PI-SceI | ATCTATGTCGGGTGCGGAGAAAGAGGTAAT(-15/-19) | NEBuffer™ PI-SceI + BSA | 10 | 10 | 10 | 10 | 65°C | 37°C | B |
PleI | GAGTC(4/5) | CutSmart® Buffer | 25 | 50 | 25 | 100 | 65°C | 37°C | A |
PluTI | GGCGC/C | CutSmart® Buffer | 100 | 25 | 10 | 100 | 65°C | 37°C | A |
PmeI | GTTT/AAAC | CutSmart® Buffer | 10 | 50 | 10 | 100 | 65°C | 37°C | A |
PmlI | CAC/GTG | CutSmart® Buffer | 100 | 50 | 10 | 100 | 65°C | 37°C | A |
PpuMI | RG/GWCCY | CutSmart® Buffer | 10 | 10 | 10 | 100 | No | 37°C | B |
PshAI | GACNN/NNGTC | CutSmart® Buffer | 25 | 50 | 10 | 100 | 65°C | 37°C | A |
PsiI | TTA/TAA | CutSmart® Buffer | 10 | 100 | 10 | 100 | 65°C | 37°C | B |
PspGI | /CCWGG | CutSmart® Buffer | 25 | 100 | 50 | 100 | No | 75°C | A |
PspOMI | G/GGCCC | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 37°C | B |
PspXI | VC/TCGAGB | CutSmart® Buffer | 10 | 100 | 25 | 100 | No | 37°C | B |
PstI § | CTGCA/G | NEBuffer™ 3.1 | 75 | 75 | 100 | 50* | 80°C | 37°C | C |
PstI-HF® | CTGCA/G | CutSmart® Buffer | 10 | 75 | 50 | 100 | No | 37°C | C |
PvuI § | CGAT/CG | NEBuffer™ 3.1 | 10 | 25 | 100 | 10 | No | 37°C | B |
PvuI-HF® | CGAT/CG | CutSmart® Buffer | 25 | 100 | 100 | 100 | No | 37°C | B |
PvuII § | CAG/CTG | NEBuffer™ 3.1 | 50 | 100 | 100 | 100* | No | 37°C | B |
PvuII-HF® | CAG/CTG | CutSmart® Buffer | 10 | 10 | 10 | 100 | No | 37°C | B |
RsaI | GT/AC | CutSmart® Buffer | 25 | 50 | 10 | 100 | No | 37°C | A |
RsrII | CG/GWCCG | CutSmart® Buffer | 25 | 75 | 10 | 100 | 65°C | 37°C | C |
SacI § | GAGCT/C | NEBuffer™ 1.1 | 100 | 50 | 10 | 100 | 65°C | 37°C | A |
SacI-HF® | GAGCT/C | CutSmart® Buffer | 10 | 50 | 10 | 100 | 65°C | 37°C | A |
SacII | CCGC/GG | CutSmart® Buffer | 10 | 100 | 10 | 100 | 65°C | 37°C | A |
SalI § | G/TCGAC | NEBuffer™ 3.1 | 10 | 10 | 100 | 10 | 65°C | 37°C | A |
SalI-HF® | G/TCGAC | CutSmart® Buffer | 10 | 100 | 100 | 100 | 65°C | 37°C | A |
SapI | GCTCTTC(1/4) | CutSmart® Buffer | 75 | 50 | 10 | 100 | 65°C | 37°C | B |
Sau3AI | /GATC | NEBuffer™ 1.1 | 100 | 50 | 10 | 100 | 65°C | 37°C | A |
Sau96I | G/GNCC | CutSmart® Buffer | 50 | 100 | 100 | 100 | 65°C | 37°C | A |
SbfI § | CCTGCA/GG | CutSmart® Buffer | 50 | 25 | 10 | 100 | 80°C | 37°C | A |
SbfI-HF® | CCTGCA/GG | CutSmart® Buffer | 50 | 25 | 10 | 100 | 80°C | 37°C | B |
ScaI-HF® | AGT/ACT | CutSmart® Buffer | 100 | 100 | 10 | 100 | 80°C | 37°C | B |
ScrFI | CC/NGG | CutSmart® Buffer | 100 | 100 | 100 | 100 | 65°C | 37°C | C |
SexAI | A/CCWGGT | CutSmart® Buffer | 100 | 75 | 50 | 100 | 65°C | 37°C | A |
SfaNI | GCATC(5/9) | NEBuffer™ 3.1 | 10 | 75 | 100 | 25 | 65°C | 37°C | B |
SfcI | C/TRYAG | CutSmart® Buffer | 75 | 50 | 25 | 100 | 65°C | 37°C | B |
SfiI | GGCCNNNN/NGGCC | CutSmart® Buffer | 25 | 100 | 50 | 100 | No | 50°C | C |
SfoI | GGC/GCC | CutSmart® Buffer | 50 | 100 | 100 | 100 | No | 37°C | B |
SgrAI | CR/CCGGYG | CutSmart® Buffer | 100 | 100 | 10 | 100 | 65°C | 37°C | A |
SmaI | CCC/GGG | CutSmart® Buffer | 10 | 10 | 10 | 100 | 65°C | 25°C | B |
SmlI | C/TYRAG | CutSmart® Buffer | 25 | 75 | 25 | 100 | No | 55°C | A |
SnaBI | TAC/GTA | CutSmart® Buffer | 50* | 50 | 10 | 100 | 80°C | 37°C | A |
SpeI § | A/CTAGT | CutSmart® Buffer | 75 | 100 | 25 | 100 | 80°C | 37°C | C |
SpeI-HF® | A/CTAGT | CutSmart® Buffer | 25 | 50 | 10 | 100 | 80°C | 37°C | C |
SphI § | GCATG/C | NEBuffer™ 2.1 | 100 | 100 | 50 | 100 | 65°C | 37°C | B |
SphI-HF® | GCATG/C | CutSmart® Buffer | 50 | 25 | 10 | 100 | 65°C | 37°C | B |
SrfI | GCCC/GGGC | CutSmart® Buffer | 10 | 50 | 0 | 100 | 65°C | 37°C | B |
SspI § | AAT/ATT | NEBuffer™ SspI | 50 | 100 | 50 | 50 | 65°C | 37°C | C |
SspI-HF® | AAT/ATT | CutSmart® Buffer | 25 | 100 | 10 | 100 | 65°C | 37°C | B |
StuI | AGG/CCT | CutSmart® Buffer | 50 | 100 | 50 | 100 | No | 37°C | A |
StyD4I | /CCNGG | CutSmart® Buffer | 10 | 100 | 100 | 100 | 65°C | 37°C | B |
StyI § | C/CWWGG | NEBuffer™ 3.1 | 10 | 25 | 100 | 10 | 65°C | 37°C | A |
StyI-HF® | C/CWWGG | CutSmart® Buffer | 25 | 100 | 25 | 100 | 65°C | 37°C | A |
SwaI | ATTT/AAAT | NEBuffer™ 3.1 | 10 | 10 | 100 | 10 | 65°C | 25°C | B |
TaqI-v2 | T/CGA | CutSmart® Buffer | 50 | 75 | 100 | 100 | 80°C | 65°C | B |
TfiI | G/AWTC | CutSmart® Buffer | 50 | 100 | 100 | 100 | No | 65°C | C |
TseI | G/CWGC | CutSmart® Buffer | 75 | 100 | 100 | 100 | No | 65°C | B |
Tsp45I | /GTSAC | CutSmart® Buffer | 100 | 50 | 10 | 100 | No | 65°C | A |
TspMI | C/CCGGG | CutSmart® Buffer | 50* | 75* | 50* | 100 | No | 75°C | B |
TspRI | NNCASTGNN/ | CutSmart® Buffer | 25 | 50 | 25 | 100 | No | 65°C | B |
Tth111I | GACN/NNGTC | CutSmart® Buffer | 25 | 100 | 25 | 100 | No | 65°C | B |
XbaI | T/CTAGA | CutSmart® Buffer | 10 | 100 | 75 | 100 | 65°C | 37°C | A |
XcmI | CCANNNNN/NNNNTGG | NEBuffer™ 2.1 | 10 | 100 | 25 | 100 | 65°C | 37°C | C |
XhoI | C/TCGAG | CutSmart® Buffer | 75 | 100 | 100 | 100 | 65°C | 37°C | A |
XmaI | C/CCGGG | CutSmart® Buffer | 25 | 50 | 10 | 100 | 65°C | 37°C | A |
XmnI | GAANN/NNTTC | CutSmart® Buffer | 50 | 75 | 10 | 100 | 65°C | 37°C | A |
ZraI | GAC/GTC | CutSmart® Buffer | 100 | 25 | 10 | 100 | 80°C | 37°C | B |
§ An HF version of this enzyme is available.
The values listed in this table are approximate. They were obtained using each enzyme's specific unit assay substrate DNA.
Ligation and Recutting Notes
a. Ligation is less than 10%.
b. Ligation is 25% -75%.
c. Recutting after ligation is less than 5%.
d. Recutting after ligation is 50% -75%.
e. Ligation and recutting after ligation is not applicable since the enzyme is either a nicking enzyme, is affected by methylation, or the recognition sequence contains variable sequences.
Star Activity Notes
1. Star Activity may result from extended digestion, high enzyme concentration or a glycerol concentration of > 5%.
2. Star Activity may result from extended digestion.
3. Star Activity may result from a glycerol concentration of > 5%.
*. May exhibit star activity in this buffer.
Simplifique a configuração da reação e a dupla digestão com o tampão CutSmart!
Mais de 215 enzimas de restrição são 100% ativas em um único tampão, o CutSmart Buffer, facilitando significativamente a configuração de suas reações de digestão dupla.
Como o CutSmart Buffer inclui BSA, também existem menos etapas de pipetagens e tubos para se preocupar.
Além disso, muitas enzimas modificadoras de DNA são 100% ativas no CutSmart Buffer, eliminando a necessidade de purificação subsequente.