碧云天生产的10X CutEZ™ Buffer是一种能让超过200种限制性内切酶的活性达到100%的缓冲液。使用CutEZ™ Buffer,可以免去内切酶酶切时选择buffer的麻烦,特别是在双酶切时会变得更加简单易行。此外,许多DNA修饰酶在CutEZ™ Buffer中也保留了100%酶活性,这样在酶切后进行DNA修饰酶处理就无需进行DNA纯化了。
各种内切酶和修饰酶在1X CutEZ™ buffer中的活性,请参考附录1和附录2。
不同来源的限制性内切酶对于CutEZ™ Buffer的兼容性可能略有不同。
1X CutEZ™ Buffer的组分为50 mM Potassium acetate, 20 mM Tris-acetate, 10 mM Magnesium acetate, 100 μg/ml BSA, pH 7.9 @ 25℃,与NEB公司的CutSmart™ Buffer完全一致。
一个包装的本产品,如果用于20μl的酶切反应体系,共可以用于2500个酶切反应。
包装清单:
产品编号 | 产品名称 | 包装 |
D6018 | 10X CutEZ™ Buffer | 1ml×5 |
— | 说明书 | 1份 |
保存条件:
-20℃保存。
注意事项:
使用前,反应缓冲液应彻底融化并混匀。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。
使用说明:
1.根据酶切反应体系加入总体积1/10的10X CutEZ™ Buffer,使其终浓度为1X,待所有组分都加入完毕,混匀后即可置于适当的温度中进行酶切反应。
附录1:
常用限制性内切酶在CutEZ™ Buffer中的活性和效能表如下:
Enzyme | Sequence | % Activity in CutEZ™ | Heat Inac. | Incu. Temp. | Dam | Dcm | CpG | Unit Substrate |
AatII | GACGT/C | 100 | 80℃ | 37℃ | ● | ● | ■ | λ DNA |
Acc65I | G/GTACC | 25 | 65℃ | 37℃ | ● | □ scol | □ scol | pBC4 DNA |
AccI | GT/MKAC | 100 | 80℃ | 37℃ | ● | ● | □ ol | λ DNA |
AciI | CCGC(-3/-1) | 100 | 65℃ | 37℃ | ● | ● | ■ | λ DNA |
AclI | AA/CGTT | 100 | No | 37℃ | ● | ● | ■ | λ DNA |
AcuI | CTGAAG(16/14) | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
AfeI | AGC/GCT | 100 | 65℃ | 37℃ | ● | ● | ■ | pXba DNA |
AflII | C/TTAAG | 100 | 65℃ | 37℃ | ● | ● | ● | ΦX174 RF I DNA |
AflIII | A/CRYGT | 50 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
AgeI | A/CCGGT | 75 | 65℃ | 37℃ | ● | ● | ■ | λ DNA |
AhdI | GACNNN/NNGTC | 100 | 65℃ | 37℃ | ● | ● | ◊ scol | λ DNA |
AleI | CACNN/NNGTG | 100 | 80℃ | 37℃ | ● | ● | ◊ scol | λ DNA |
AluI | AG/CT | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
AlwI | GGATC(4/5) | 100 | No | 37℃ | ■ | ● | ● | λ DNA (dam-) |
AlwNI | CAGNNN/CTG | 100 | 80℃ | 37℃ | ● | □ ol | ● | λ DNA |
ApaI | GGGCC/C | 100 | 65℃ | 25℃ | ● | □ ol | □ ol | pXba DNA |
ApaLI | G/TGCAC | 100 | No | 37℃ | ● | ● | □ ol | λ DNA/HindIII |
ApeKI | G/CWGC | 10 | No | 75℃ | ● | ● | λ DNA | |
ApoI | R/AATTY | 75 | 80℃ | 50℃ | ● | ● | ● | λ DNA |
AscI | GG/CGCGCC | 100 | 80℃ | 37℃ | ● | ● | ■ | λ DNA |
AseI | AT/TAAT | 10 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
AsiSI | GCGAT/CGC | 100 | 80℃ | 37℃ | ● | ● | ■ | pXba/XhoI |
AvaI | C/YCGRG | 100 | 80℃ | 37℃ | ● | ● | ■ | λ DNA |
AvaII | G/GWCC | 100 | 80℃ | 37℃ | ● | □ ol | □ ol | λ DNA |
AvrII | C/CTAGG | 100 | No | 37℃ | ● | ● | ● | λ DNA/HindIII |
BaeI | (10/15)ACNNNNGTAYC(12/7) | 100 | 65℃ | 25℃ | ● | ● | □ scol | pXba DNA |
BamHI | G/GATCC | 100* | No | 37℃ | ● | ● | ● | λ DNA |
BanI | G/GYRCC | 100 | 65℃ | 37℃ | ● | □ scol | □ scol | λ DNA |
BanII | GRGCY/C | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
BbsI | GAAGAC(2/6) | 75 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
BbvCI | CCTCAGC(-5/-2) | 100 | No | 37℃ | ● | ● | ◊ ol | λ DNA |
BbvI | GCAGC(8/12) | 100 | 65℃ | 37℃ | ● | ● | ● | pBR322 (dcm+) DNA |
BccI | CCATC(4/5) | 100 | 65℃ | 37℃ | ● | ● | ● | pXba DNA |
BceAI | ACGGC(12/14) | 100* | 65℃ | 37℃ | ● | ● | ■ | pBR322 (dcm+) DNA |
BcgI | (10/12)CGANNNNNNTGC(12/10) | 50* | 65℃ | 37℃ | ◊ ol | ● | □ scol | λ DNA |
BciVI | GTATCC(6/5) | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
BclI | T/GATCA | 75 | No | 50℃ | ■ | ● | ● | λ DNA (dam-) |
BfaI | C/TAG | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
BfuAI | ACCTGC(4/8) | 10 | 65℃ | 50℃ | ● | ● | ◊ ol | λ DNA |
BglI | GCCNNNN/NGGC | 10 | 65℃ | 37℃ | ● | ● | □ scol | λ DNA |
BglII | A/GATCT | 10 | No | 37℃ | ● | ● | ● | λ DNA |
BlpI | GC/TNAGC | 100 | No | 37℃ | ● | ● | ● | λ DNA |
BmgBI | CACGTC(-3/-3) | 10 | 65℃ | 37℃ | ● | ● | ■ | λ DNA |
BmrI | ACTGGG(5/4) | 100* | 65℃ | 37℃ | ● | ● | ● | λ DNA/HindIII |
BmtI | GCTAG/C | 100 | 65℃ | 37℃ | ● | ● | ● | pXba DNA |
BpmI | CTGGAG(16/14) | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
Bpu10I | CCTNAGC(-5/-2) | 25 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
BpuEI | CTTGAG(16/14) | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
BsaAI | YAC/GTR | 100 | No | 37℃ | ● | ● | ■ | λ DNA |
BsaBI | GATNN/NNATC | 100 | 80℃ | 60℃ | □ ol | ● | □ scol | λ DNA (dam-) |
BsaHI | GR/CGYC | 100 | 80℃ | 37℃ | ● | □ scol | ■ | λ DNA |
BsaI | GGTCTC(1/5) | 100 | 65℃ | 37℃ | ● | ◊ scol | □ scol | pXba DNA |
BsaJI | C/CNNGG | 100 | 80℃ | 60℃ | ● | ● | ● | λ DNA |
BsaWI | W/CCGGW | 100 | 80℃ | 60℃ | ● | ● | ● | λ DNA |
BsaXI | (9/12)ACNNNNNCTCC(10/7) | 100 | No | 37℃ | ● | ● | ● | λ DNA |
BseRI | GAGGAG(10/8) | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
BseYI | CCCAGC(-5/-1) | 50 | 80℃ | 37℃ | ● | ● | □ ol | λ DNA |
BsgI | GTGCAG(16/14) | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
BsiEI | CGRY/CG | 100 | No | 60℃ | ● | ● | ■ | λ DNA |
BsiHKAI | GWGCW/C | 100 | No | 65℃ | ● | ● | ● | λ DNA |
BsiWI | C/GTACG | 25 | 65℃ | 55℃ | ● | ● | ■ | ΦX174 DNA |
BslI | CCNNNNN/NNGG | 100 | No | 55℃ | ● | □ scol | □ scol | λ DNA |
BsmAI | GTCTC(1/5) | 100 | No | 55℃ | ● | ● | □ scol | λ DNA |
BsmBI | CGTCTC(1/5) | 25 | 80℃ | 55℃ | ● | ● | ■ | λ DNA |
BsmFI | GGGAC(10/14) | 100 | 80℃ | 65℃ | ● | □ ol | □ ol | pBR322 (dcm+) DNA |
BsmI | GAATGC(1/-1) | 100 | 80℃ | 65℃ | ● | ● | ● | λ DNA |
BsoBI | C/YCGRG | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
Bsp1286I | GDGCH/C | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
BspCNI | CTCAG(9/7) | 100 | 80℃ | 25℃ | ● | ● | ● | λ DNA |
BspDI | AT/CGAT | 100 | 80℃ | 37℃ | □ ol | ● | ■ | λ DNA |
BspEI | T/CCGGA | 10 | 80℃ | 37℃ | □ ol | ● | ♦ | λ DNA (dam-) |
BspHI | T/CATGA | 100 | 80℃ | 37℃ | ◊ ol | ● | ● | λ DNA |
BspMI | ACCTGC(4/8) | 10 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
BspQI | GCTCTTC(1/4) | 100 | 80℃ | 50℃ | ● | ● | ● | λ DNA |
BsrBI | CCGCTC(-3/-3) | 100 | 80℃ | 37℃ | ● | ● | □ scol | λ DNA |
BsrDI | GCAATG(2/0) | 25 | 80℃ | 65℃ | ● | ● | ● | λ DNA |
BsrFI | R/CCGGY | 100 | No | 37℃ | ● | ● | ■ | pBR322 (dcm+) DNA |
BsrGI | T/GTACA | 25 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
BsrI | ACTGG(1/-1) | 10 | 80℃ | 65℃ | ● | ● | ● | ΦX174 RF I DNA |
BssHII | G/CGCGC | 100 | 65℃ | 50℃ | ● | ● | ■ | λ DNA |
BssKI | /CCNGG | 100 | 80℃ | 60℃ | ● | □ ol | □ ol | λ DNA |
BssSI | CACGAG(-5/-1) | 50 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
BstAPI | GCANNNN/NTGC | 100 | 80℃ | 60℃ | ● | ● | □ scol | λ DNA |
BstBI | TT/CGAA | 100 | No | 65℃ | ● | ● | ■ | λ DNA |
BstEII | G/GTNACC | 75* | No | 60℃ | ● | ● | ● | λ DNA |
BstNI | CC/WGG | 75 | No | 60℃ | ● | ● | ● | λ DNA |
BstUI | CG/CG | 100 | No | 60℃ | ● | ● | ■ | λ DNA |
BstXI | CCANNNNN/NTGG | 25 | 80℃ | 37℃ | ● | □ scol | ● | λ DNA |
BstYI | R/GATCY | 100 | No | 60℃ | ● | ● | ● | λ DNA |
BstZ17I | GTA/TAC | 100 | No | 37℃ | ● | ● | □ scol | λ DNA |
Bsu36I | CC/TNAGG | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA/HindIII |
BtgI | C/CRYGG | 100 | 80℃ | 37℃ | ● | ● | ● | pBR322 (dcm+) DNA |
BtgZI | GCGATG(10/14) | 100 | 80℃ | 60℃ | ● | ● | ♦ | λ DNA |
BtsCI | GGATG(2/0) | 100 | 80℃ | 50℃ | ● | ● | ● | λ DNA |
BtsI | GCAGTG(2/0) | 100 | 80℃ | 55℃ | ● | ● | ● | λ DNA |
Cac8I | GCN/NGC | 100 | 65℃ | 37℃ | ● | ● | □ scol | λ DNA |
ClaI | AT/CGAT | 100 | 65℃ | 37℃ | □ ol | ● | ■ | λ DNA (dam-) |
CspCI | (11/13)CAANNNNNGTGG(12/10) | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
CviAII | C/ATG | 100 | 65℃ | 25℃ | ● | ● | ● | λ DNA |
CviKI-1 | RG/CY | 100 | No | 37℃ | ● | ● | ● | pBR322 (dcm+) DNA |
CviQI | G/TAC | 75* | No | 25℃ | ● | ● | ● | λ DNA |
DdeI | C/TNAG | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
DpnI | GA/TC | 100 | 80℃ | 37℃ | ● | ● | □ ol | pBR322 DNA |
DpnII | /GATC | 25 | 65℃ | 37℃ | ■ | ● | ● | λ DNA (dam-) |
DraI | TTT/AAA | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
DraIII | CACNNN/GTG | 25 | 65℃ | 37℃ | ● | ● | ◊ ol | λ DNA |
DrdI | GACNNNN/NNGTC | 100 | 65℃ | 37℃ | ● | ● | □ scol | pUC19 DNA |
EaeI | Y/GGCCR | 100 | 65℃ | 37℃ | ● | □ ol | □ ol | λ DNA |
EagI | C/GGCCG | 10 | 65℃ | 37℃ | ● | ● | ■ | pXba DNA |
EarI | CTCTTC(1/4) | 100 | 65℃ | 37℃ | ● | ● | ◊ ol | λ DNA |
EciI | GGCGGA(11/9) | 100 | 65℃ | 37℃ | ● | ● | □ scol | λ DNA |
EcoNI | CCTNN/NNNAGG | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
EcoO109I | RG/GNCCY | 100 | 65℃ | 37℃ | ● | □ ol | ● | λ DNA/HindIII |
EcoP15I | CAGCAG(25/27) | 100 | 65℃ | 37℃ | ● | ● | ● | pUC19 DNA |
EcoRI | G/AATTC | 50* | 65℃ | 37℃ | ● | ● | □ scol | λ DNA |
EcoRV | GAT/ATC | 10 | 80℃ | 37℃ | ● | ● | ◊ scol | λ DNA |
FatI | /CATG | 50 | 80℃ | 55℃ | ● | ● | ● | pUC19 DNA |
FauI | CCCGC(4/6) | 100 | 65℃ | 55℃ | ● | ● | ■ | λ DNA |
Fnu4HI | GC/NGC | 100 | No | 37℃ | ● | ● | □ ol | λ DNA |
FokI | GGATG(9/13) | 100 | 65℃ | 37℃ | ● | ◊ ol | ◊ ol | λ DNA |
FseI | GGCCGG/CC | 100 | 65℃ | 37℃ | ● | ◊ scol | ■ | Adenovirus-2 DNA |
FspI | TGC/GCA | 100 | No | 37℃ | ● | ● | ■ | λ DNA |
HaeII | RGCGC/Y | 100 | 80℃ | 37℃ | ● | ● | ■ | λ DNA |
HaeIII | GG/CC | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
HgaI | GACGC(5/10) | 100 | 65℃ | 37℃ | ● | ● | ■ | ΦX174 DNA |
HhaI | GCG/C | 100 | 65℃ | 37℃ | ● | ● | ■ | λ DNA |
HincII | GTY/RAC | 100 | 65℃ | 37℃ | ● | ● | □ scol | λ DNA |
HindIII | A/AGCTT | 50 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
HinfI | G/ANTC | 100 | 80℃ | 37℃ | ● | ● | □ scol | λ DNA |
HinP1I | G/CGC | 100 | 65℃ | 37℃ | ● | ● | ■ | λ DNA |
HpaI | GTT/AAC | 100 | No | 37℃ | ● | ● | □ scol | λ DNA |
HpaII | C/CGG | 100 | 80℃ | 37℃ | ● | ● | ■ | λ DNA |
HphI | GGTGA(8/7) | 100 | 65℃ | 37℃ | ■ | ■ | ● | λ DNA |
Hpy188I | TCN/GA | 100 | 65℃ | 37℃ | □ scol | ● | ● | pBR322 (dcm+) |
Hpy99I | CGWCG/ | 100 | 65℃ | 37℃ | ● | ● | ■ | λ DNA |
HpyAV | CCTTC(6/5) | 100 | 65℃ | 37℃ | ● | ● | ◊ ol | λ DNA |
HpyCH4III | ACN/GT | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
HpyCH4I | A/CGT | 100 | 65℃ | 37℃ | ● | ● | ■ | pUC19 DNA |
HpyCH4V | TG/CA | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
KasI | G/GCGCC | 100 | 65℃ | 37℃ | ● | ● | ■ | pBR322 DNA |
KpnI | GGTAC/C | 50* | No | 37℃ | ● | ● | ● | pXba DNA |
MboI | /GATC | 100 | 65℃ | 37℃ | ■ | ● | ◊ ol | λ DNA (dam-) |
MboII | GAAGA(8/7) | 100 | 65℃ | 37℃ | □ ol | ● | ● | λ DNA (dam-) |
MfeI | C/AATTG | 100 | No | 37℃ | ● | ● | ● | λ DNA |
MluI | A/CGCGT | 25 | 80℃ | 37℃ | ● | ● | ■ | λ DNA |
MlyI | GAGTC(5/5) | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
MmeI | TCCRAC(20/18) | 100 | 65℃ | 37℃ | ● | ● | □ ol | ΦX174 DNA |
MnlI | CCTC(7/6) | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
MscI | TGG/CCA | 100 | 80℃ | 37℃ | ● | □ ol | ● | λ DNA |
MseI | T/TAA | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
MslI | CAYNN/NNRTG | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
MspA1I | CMG/CKG | 100 | 65℃ | 37℃ | ● | ● | □ ol | λ DNA |
MspI | C/CGG | 100 | No | 37℃ | ● | ● | ● | λ DNA |
MwoI | GCNNNNN/NNGC | 100 | No | 60℃ | ● | ● | □ scol | λ DNA |
NaeI | GCC/GGC | 100 | No | 37℃ | ● | ● | ■ | pXba DNA |
NarI | GG/CGCC | 100 | 65℃ | 37℃ | ● | ● | ■ | pXba DNA |
NciI | CC/SGG | 100 | No | 37℃ | ● | ● | ◊ ol | λ DNA |
NcoI | C/CATGG | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
NdeI | CA/TATG | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
NgoMIV | G/CCGGC | 100 | No | 37℃ | ● | ● | ■ | Adenovirus-2 DNA |
NheI | G/CTAGC | 100 | 65℃ | 37℃ | ● | ● | □ scol | λ DNA/HindIII |
NlaIII | CATG/ | 100 | 65℃ | 37℃ | ● | ● | ● | ΦX174 DNA |
NlaIV | GGN/NCC | 100 | 65℃ | 37℃ | ● | □ ol | □ ol | pBR322 DNA |
NmeAIII | GCCGAG(21/19) | 100 | 65℃ | 37℃ | ● | ● | ● | ΦX174 RF I DNA |
NotI | GC/GGCCGC | 25 | 65℃ | 37℃ | ● | ● | ■ | pBC4 DNA |
NruI | TCG/CGA | 10 | No | 37℃ | □ ol | ● | ■ | λ DNA |
NsiI | ATGCA/T | 25 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
NspI | RCATG/Y | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
PacI | TTAAT/TAA | 100 | 65℃ | 37℃ | ● | ● | ● | pNEB193 DNA |
PaeR7I | C/TCGAG | 100 | No | 37℃ | ● | ● | ■ | λ DNA/HindIII |
PciI | A/CATGT | 50* | 80℃ | 37℃ | ● | ● | ● | pXba DNA |
PflFI | GACN/NNGTC | 100 | 65℃ | 37℃ | ● | ● | ● | pBC4 DNA |
PflMI | CCANNNN/NTGG | 50 | 65℃ | 37℃ | ● | □ ol | ● | λ DNA |
PhoI | GG/CC | 100 | No | 37℃ | ● | ◊ scol | ◊ scol | λ DNA |
PleI | GAGTC(4/5) | 100 | 65℃ | 37℃ | ● | ● | □ scol | λ DNA |
PmeI | GTTT/AAAC | 100 | 65℃ | 37℃ | ● | ● | □ scol | λ DNA |
PmlI | CAC/GTG | 100 | 65℃ | 37℃ | ● | ● | ■ | λ DNA/HindIII |
PpuMI | RG/GWCCY | 100 | No | 37℃ | ● | □ ol | ● | λ DNA/HindIII |
PshAI | GACNN/NNGTC | 100 | 65℃ | 37℃ | ● | ● | □ scol | λ DNA |
PsiI | TTA/TAA | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
PspGI | /CCWGG | 100 | No | 75℃ | ● | ■ | ● | T7 DNA |
PspOMI | G/GGCCC | 100 | 65℃ | 37℃ | ● | ◊ scol | □ ol | pXba DNA |
PspXI | VC/TCGAGB | 100 | No | 37℃ | ● | ● | ♦ | λ DNA/HindIII |
PstI | CTGCA/G | 50* | 80℃ | 37℃ | ● | ● | ● | λ DNA |
PvuI | CGAT/CG | 10 | No | 37℃ | ● | ● | ■ | pXba DNA |
PvuII | CAG/CTG | 100* | No | 37℃ | ● | ● | ● | λ DNA |
RsaI | GT/AC | 100 | No | 37℃ | ● | ● | □ scol | λ DNA |
RsrII | CG/GWCCG | 100 | 65℃ | 37℃ | ● | ● | ■ | λ DNA |
SacI | GAGCT/C | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA/HindIII |
SacII | CCGC/GG | 100 | 65℃ | 37℃ | ● | ● | ■ | pXba DNA |
SalI | G/TCGAC | 10 | 65℃ | 37℃ | ● | ● | ■ | λ DNA/HindIII |
SapI | GCTCTTC(1/4) | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
Sau3AI | /GATC | 100 | 65℃ | 37℃ | ● | ● | □ ol | λ DNA |
Sau96I | G/GNCC | 100 | 65℃ | 37℃ | ● | □ ol | □ ol | λ DNA |
SbfI | CCTGCA/GG | 100 | 80℃ | 37℃ | ● | ● | ● | λ DNA |
ScaI | AGT/ACT | N/R | 80℃ | 37℃ | ● | ● | ● | λ DNA |
ScrFI | CC/NGG | 100 | 65℃ | 37℃ | ● | □ ol | □ ol | λ DNA |
SexAI | A/CCWGGT | 100 | 65℃ | 37℃ | ● | ■ | ● | pBC4 DNA (dcm-) |
SfaNI | GCATC(5/9) | 25 | 65℃ | 37℃ | ● | ● | ◊ scol | ΦX174 RF I DNA |
SfcI | C/TRYAG | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
SfiI | GGCCNNNN/NGGCC | 100 | No | 50℃ | ● | ◊ ol | □ scol | Adenovirus-2 DNA |
SfoI | GGC/GCC | 100 | No | 37℃ | ● | □ scol | ■ | λ DNA/HindIII |
SgrAI | CR/CCGGYG | 100 | 65℃ | 37℃ | ● | ● | ■ | λ DNA |
SmaI | CCC/GGG | 100 | 65℃ | 25℃ | ● | ● | ■ | λ DNA |
SmlI | C/TYRAG | 100 | No | 55℃ | ● | ● | ● | λ DNA |
SnaBI | TAC/GTA | 100 | 80℃ | 37℃ | ● | ● | ■ | T7 DNA |
SpeI | A/CTAGT | 100 | 80℃ | 37℃ | ● | ● | ● | Adenovirus-2 DNA |
SphI | GCATG/C | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
SspI | AAT/ATT | 50 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
StuI | AGG/CCT | 100 | No | 37℃ | ● | □ ol | ● | λ DNA |
StyD4I | /CCNGG | 100 | 65℃ | 37℃ | ● | □ ol | ◊ ol | λ DNA |
StyI | C/CWWGG | 10 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
SwaI | ATTT/AAAT | 10 | 65℃ | 25℃ | ● | ● | ● | M13mp19 RF I DNA |
TaqαI | T/CGA | 100 | 80℃ | 65℃ | □ ol | ● | ● | λ DNA |
TfiI | G/AWTC | 100 | No | 65℃ | ● | ● | □ scol | λ DNA |
TliI | C/TCGAG | 50 | No | 75℃ | ● | ● | ♦ | λ DNA/HindIII |
TseI | G/CWGC | 100 | No | 65℃ | ● | ● | □ scol | λ DNA |
Tsp45I | /GTSAC | 100 | No | 65℃ | ● | ● | ● | λ DNA |
Tsp509I | /AATT | N/R | No | 65℃ | ● | ● | ● | λ DNA |
TspMI | C/CCGGG | 100 | No | 75℃ | ● | ● | ■ | pUCAdeno |
TspRI | NNCASTGNN/ | 100 | No | 65℃ | ● | ● | ● | λ DNA |
Tth111I | GACN/NNGTC | 100 | No | 65℃ | ● | ● | ● | pBC4 DNA |
XbaI | T/CTAGA | 100 | 65℃ | 37℃ | □ ol | ● | ● | λ DNA(dam-)/Hind III |
XcmI | CCANNNNN/NNNNTGG | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
XhoI | C/TCGAG | 100 | 65℃ | 37℃ | ● | ● | ♦ | λ DNA/HindIII |
XmaI | C/CCGGG | 100 | 65℃ | 37℃ | ● | ● | ♦ | Adenovirus-2 DNA |
XmnI | GAANN/NNTTC | 100 | 65℃ | 37℃ | ● | ● | ● | λ DNA |
ZraI | GAC/GTC | 100 | 80℃ | 37℃ | ● | ● | ■ | λ DNA |
注:*,在此buffer中可能产生星号活性。λ DNA/HindIII:λ DNA digested with HindIII.
● 不敏感
■ 完全阻断
□ ol 重叠性阻断
□ scol 某些重叠组合后被阻断
♦ 部分阻断
◊ ol 重叠性部分阻断
◊ scol 某些重叠组合后被部分阻断
附录2:
DNA修饰酶在CutEZ™ Buffer中的活性表格如下:
Enzyme | % Activity in CutEZ™ | Required Supplements |
Alkaline Phosphatase(CIP) | 100 | - |
Antarctic Phosphatase | 100 | Zn2+ |
Bst DNA Polymerase | 100 | - |
CpG Methyltransferase(M.SssI) | 100 | - |
DNA Polymerase Ⅰ | 100 | - |
DNA PolymeraseⅠ, Large(klenow)Fragment | 100 | - |
DNA Polymerase Klenow Exo- | 100 | - |
DNaseⅠ(RNase-free) | 100 | Ca2+ |
E.coli DNA Ligase | 100 | NAD |
Endonuclease Ⅲ(Nth), recombinant | 100 | - |
Endonuclease VIII | 100 | - |
Exonuclease Ⅲ | 100 | - |
GpC Methyltransferase(M.CviPI) | 0-50 | DTT |
McrBC | 100 | - |
Micrococcal Nuclease | 100 | - |
Nuclease BAL-31 | 100 | - |
phi29 DNA Polymerase | 100 | - |
RecJf | 100 | - |
T3 DNA Ligase | 100 | ATP+PEG |
T4 DNA Ligase | 100 | ATP |
T4 DNA Polymerase | 100 | - |
T4 Phage β-glucosyltransferase(T4-BGT) | 100 | - |
T4 Polynucleotide Kinase | 100 | ATP |
T4 PNK(3' phosphatase minus) | 100 | ATP |
T7 DNA Ligase | 100 | ATP+PEG |
T7 DNA Polymerase(unmodified) | 100 | - |
T7 Exonuclease | 100 | - |
USER Enzyme, recombinant | 100 | - |