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限制酶在各种NEBuffer 中的活性 (NEBuffer Activity Chart for Restriction Enzymes)
对应每一种限制性内切酶,New England Biolabs 均提供彩色盖子的10X NEBuffer,以保证100%活性。下面表格中列出每一种酶及其随酶提供的最适宜的NEBuffer。同时也列出每种酶在四种不同的NEBuffer中的大致活性,以帮助选择双酶切时最适宜的环境。除非特别指出,温育温度均为37°C。
注意事项:表格中所列出的酶活性是近似值,计算酶活性时所采用的底物与计算该酶的单位时所采用的底物相同。
缓冲液在使用前应充分解冻,并混合均匀。
| Enzyme |
Supplied NEBuffer |
% Activity in NEBuffers |
| 1 |
2 |
3 |
4 |
| Aat II |
4 |
0 |
50 |
50 |
100 |
| Acc I |
4 |
50 |
50 |
10 |
100 |
| Acc65 I |
3 + BSA |
10 |
75 |
100 |
25 |
| Aci I |
3 |
25 |
50 |
100 |
50 |
| Acl I |
4 + BSA |
10 |
10 |
0 |
100 |
| Afe I * |
SE-Y |
25 |
50 |
25 |
100 |
| Afl II |
2 + BSA |
50 |
100 |
25 |
100 |
| Afl III |
3 + BSA |
25 |
75 |
100 |
50 |
| Age I |
1 |
100 |
50 |
10 |
75 |
| Ahd I |
4 + BSA |
25 |
75 |
0 |
100 |
| Ale I |
4 |
10 |
20 |
10 |
100 |
| Alu I |
2 |
100 |
100 |
75 |
100 |
| Alw I |
4 |
50 |
100 |
10 |
100 |
| AlwN I |
4 |
50 |
100 |
50 |
100 |
| Apa I @25°C |
4 + BSA |
25 |
50 |
0 |
100 |
| ApaL I |
4 + BSA |
100 |
100 |
10 |
100 |
| Apo I @50°C |
3 + BSA |
10 |
75 |
100 |
75 |
| Asc I |
4 |
0 |
10 |
10 |
100 |
| Ase I |
3 |
NR |
75 |
dd100 |
NR |
| AsiS I |
3 + BSA |
50 |
100 |
100 |
50 |
| Ava I |
4 |
10 |
75 |
10 |
100 |
| Ava II |
4 |
50 |
75 |
10 |
100 |
| Avr II |
2 |
100 |
100 |
50 |
100 |
| Bae I @25°C |
2 + BSA + SAM |
50 |
100 |
50 |
75 |
| BamH I |
Udd + BSA |
75 |
100 |
50 |
75 |
| Ban I |
4 |
50 |
100 |
50 |
100 |
| Ban II |
4 |
100 |
10 |
50 |
100 |
| Bbs I |
2 |
100 |
100 |
25 |
75 |
| Bbv I |
2 |
100 |
100 |
25 |
75 |
| BbvC I |
4 |
50 |
100 |
10 |
100 |
| BceA I |
Udd + BSA |
25 |
NR |
NR |
75 |
| Bcg I |
U + SAM |
50 |
75 |
100 |
50 |
| BciV I |
4 |
100 |
25 |
10 |
100 |
| Bcl I @50°C |
3 |
50 |
100 |
dd100 |
75 |
| Bfa I |
4 |
75 |
50 |
10 |
100 |
| BfrB I |
4 + BSA |
10 |
25 |
10 |
100 |
| BfuA I @50°C |
3 |
0 |
75 |
100 |
10 |
| BfuC I |
4 + BSA |
100 |
50 |
25 |
100 |
| Bgl I |
3 |
50 |
75 |
100 |
50 |
| Bgl II |
3 |
10 |
75 |
100 |
10 |
| Blp I |
4 |
50 |
100 |
10 |
100 |
| Bme1580 I |
2 |
75 |
100 |
75 |
75 |
| BmgB I |
3 + BSA |
0 |
25 |
100 |
10 |
| Bmr I |
2 |
75 |
100 |
75 |
100 |
| Bmt I * |
SE-W |
25 |
100 |
10 |
50 |
| Bpm I |
3 + BSA |
75 |
100 |
100 |
100 |
| Bpu10 I* |
SE-K |
0 |
25 |
100 |
25 |
| BpuE I @25°C |
2 + SAM |
75 |
100 |
10 |
75 |
| Bsa I @50°C |
3 |
75 |
75 |
100 |
100 |
| BsaA I |
3 |
75 |
75 |
100 |
75 |
| BsaB I @60°C |
2 |
75 |
100 |
100 |
100 |
| BsaH I |
4 + BSA |
50 |
100 |
100 |
100 |
| BsaJ I @60°C |
2 |
100 |
100 |
100 |
100 |
| BsaW I @60°C |
2 + BSA |
50 |
100 |
50 |
50 |
| BsaX I |
4 |
75 |
100 |
10 |
100 |
| BseR I |
2 |
100 |
100 |
75 |
100 |
| BseY I |
3 |
10 |
50 |
100 |
50 |
| Bsg I |
4 + SAM |
50 |
75 |
50 |
100 |
| BsiE I @60°C |
2 + BSA |
50 |
100 |
10 |
100 |
| BsiHKA I @65°C |
3 + BSA |
50 |
100 |
100 |
100 |
| BsiW I @55°C |
3 |
100 |
100 |
100 |
25 |
| Bsl I @55°C |
3 |
10 |
50 |
100 |
75 |
| Bsm I @65°C |
2 |
75 |
100 |
75 |
75 |
| BsmA I @55°C |
3 |
50 |
100 |
100 |
50 |
| BsmB I @55°C |
3 |
75 |
100 |
100 |
100 |
| BsmF I @65°C |
4 + BSA |
25 |
50 |
50 |
100 |
| BsoB I @65°C |
2 |
10 |
100 |
100 |
50 |
| Bsp1286 I |
4 + BSA |
25 |
25 |
25 |
100 |
| BspCN I @25°C |
1 + BSA + SAM |
100 |
75 |
10 |
75 |
| BspD I |
4 |
25 |
75 |
50 |
100 |
| BspE I |
3 |
0 |
10 |
100 |
0 |
| BspH I |
4 |
10 |
100 |
50 |
100 |
| BspM I |
3 |
NR |
NR |
100 |
NR |
| Bsr I @65°C |
3 |
0 |
50 |
100 |
10 |
| BsrB I |
2 |
50 |
100 |
100 |
100 |
| BsrD I @60°C |
2 + BSA |
50 |
100 |
50 |
75 |
| BsrF I |
U |
25 |
100 |
75 |
75 |
| BsrG I |
2 + BSA |
25 |
100 |
10 |
100 |
| BssH II @50°C |
3 |
100 |
100 |
dd100 |
100 |
| BssK I @60°C |
3 + BSA |
0 |
50 |
100 |
50 |
| BssS I |
3 |
0 |
50 |
dd100 |
10 |
| BstAP I * @60°C |
SE-W |
25 |
25 |
100 |
25 |
| BstB I @65°C |
4 |
75 |
50 |
25 |
100 |
| BstE II @60°C |
3 |
50 |
75 |
100 |
75 |
| BstF5 I * @65°C |
SE-Y |
75 |
50 |
25 |
100 |
| BstN I @60°C |
2 + BSA |
10 |
100 |
100 |
75 |
| BstU I @60°C |
2 |
100 |
100 |
50 |
100 |
| BstX I @55°C |
3 |
25 |
100 |
100 |
50 |
| BstY I @60°C |
2 |
50 |
100 |
75 |
100 |
| BstZ17 I |
3 |
NR |
NR |
100 |
NR |
| Bsu36 I |
3 + BSA |
0 |
25 |
100 |
0 |
| Btg I |
3 + BSA |
25 |
50 |
100 |
100 |
| Bts II |
4 + BSA |
100 |
50 |
50 |
100 |
| Cac8 I |
3 |
50 |
75 |
100 |
75 |
| Cla I |
4 + BSA |
10 |
50 |
50 |
100 |
| Dde I |
3 |
75 |
100 |
100 |
75 |
| Dpn I |
4 |
100 |
100 |
75 |
100 |
| Dpn II |
U |
NR |
NR |
dd100 |
NR |
| Dra I |
4 |
75 |
75 |
50 |
100 |
| Dra III |
3 + BSA |
100 |
75 |
100 |
25 |
| Drd I |
4 |
25 |
50 |
10 |
100 |
| Eae I |
1 |
100 |
100 |
50 |
100 |
| Eag I |
3 |
10 |
25 |
100 |
10 |
| Ear I |
1 |
100 |
100 |
50 |
100 |
| Eci I |
2 + BSA |
50 |
100 |
100 |
50 |
| EcoN I |
4 |
100 |
100 |
75 |
100 |
| EcoO109 I |
4 + BSA |
100 |
100 |
75 |
100 |
| EcoR I |
Udd |
100 |
100 |
100 |
100 |
| EcoR V |
3 + BSA |
50 |
75 |
100 |
50 |
| Fat I * |
SE-G |
10 |
100 |
50 |
50 |
| Fau I * @55°C |
SE-B |
100 |
25 |
10 |
50 |
| Fnu4H I |
4 |
10 |
25 |
25 |
100 |
| Fok I |
4 |
50 |
100 |
75 |
100 |
| Fse I |
4 + BSA |
100 |
75 |
0 |
100 |
| Fsp I |
4 |
10 |
75 |
10 |
100 |
| Hae II |
4 + BSA |
75 |
100 |
50 |
100 |
| Hae III |
2 |
50 |
100 |
25 |
75 |
| Hga I |
1 |
100 |
75 |
50 |
100 |
| Hha I |
4 + BSA |
75 |
100 |
100 |
100 |
| Hinc II |
3 + BSA |
75 |
100 |
100 |
100 |
| Hind III |
2 |
50 |
100 |
10 |
50 |
| Hinf I |
2 |
75 |
100 |
75 |
75 |
| HinP1 I |
2 |
100 |
100 |
100 |
100 |
| Hpa I |
4 |
25 |
50 |
10 |
100 |
| Hpa II |
1 |
100 |
50 |
10 |
100 |
| Hph I |
4 |
NR |
75 |
0 |
100 |
| Hpy99 I |
4 + BSA |
10 |
10 |
0 |
100 |
| Hpy188 I |
4 |
50 |
25 |
10 |
100 |
| Hpy188 III |
4 + BSA |
100 |
100 |
10 |
100 |
| HpyCH4III |
4 |
100 |
50 |
25 |
100 |
| HpyCH4IV |
1 |
100 |
25 |
10 |
25 |
| HpyCH4V |
4 |
50 |
50 |
25 |
100 |
| I-Ceu I |
4 + BSA |
10 |
10 |
0 |
100 |
| I-Sce I |
U + BSA |
10 |
50 |
50 |
50 |
| Kas I |
2 + BSA |
75 |
100 |
75 |
75 |
| Kpn I |
1 + BSA |
100 |
75 |
0 |
50 |
| Mbo I |
3 |
75 |
100 |
100 |
100 |
| Mbo II |
2 |
100 |
100 |
50 |
100 |
| McrBC |
2 + BSA + GTP |
50 |
100 |
10 |
75 |
| Mfe I |
4 |
75 |
50 |
10 |
100 |
| Mlu I |
3 |
25 |
75 |
100 |
50 |
| Mly I |
4 + BSA |
50 |
50 |
25 |
100 |
| Mme I |
4 + SAM |
NR |
NR |
NR |
100 |
| Mnl I |
2 + BSA |
75 |
100 |
50 |
75 |
| Msc I |
4 |
75 |
75 |
75 |
100 |
| Mse I |
2 + BSA |
75 |
100 |
75 |
75 |
| Msl I |
2 |
100 |
100 |
25 |
100 |
| Msp I |
2 |
75 |
100 |
50 |
50 |
| MspA1 I |
4 + BSA |
50 |
100 |
50 |
100 |
| Mwo I @60°C |
U |
10 |
75 |
100 |
75 |
| Nae I |
1 |
100 |
75 |
10 |
75 |
| N.Alw I |
2 |
25 |
100 |
25 |
50 |
| Nar I |
1 |
100 |
75 |
75 |
75 |
| N.BbvC IA |
2 |
50 |
100 |
10 |
100 |
| N.BbvC IB |
2 |
50 |
100 |
10 |
100 |
| N.BstNB I @55°C |
U |
0 |
10 |
10 |
10 |
| Nci I |
4 |
100 |
25 |
10 |
100 |
| Nco I |
4 |
100 |
100 |
100 |
100 |
| Nde I |
4 |
75 |
100 |
75 |
100 |
| NgoM IV |
4 |
100 |
50 |
10 |
100 |
| Nhe I |
2 + BSA |
100 |
100 |
10 |
100 |
| Nla III |
4 + BSA |
25 |
25 |
25 |
100 |
| Nla IV |
4 + BSA |
10 |
10 |
10 |
100 |
| Not I |
3 + BSA |
0 |
75 |
100 |
50 |
| Nru I |
U |
0 |
10 |
75 |
10 |
| Nsi I |
U |
25 |
25 |
50 |
10 |
| Nsp I |
2 + BSA |
100 |
100 |
0 |
100 |
| Pac I |
1 + BSA |
100 |
75 |
10 |
100 |
| PaeR7 I |
4 |
25 |
100 |
10 |
100 |
| Pci I * |
SE-O |
25 |
75 |
100 |
50 |
| PflF I |
4 + BSA |
0 |
25 |
25 |
100 |
| PflM I |
3 + BSA |
0 |
100 |
100 |
50 |
| PI-Psp I @65°C |
U + BSA |
0 |
10 |
10 |
10 |
| PI-Sce I |
U + BSA |
0 |
0 |
0 |
0 |
| Ple I |
4 |
100 |
100 |
75 |
100 |
| Pme I |
4 + BSA |
0 |
50 |
10 |
100 |
| Pml I |
1 + BSA |
100 |
75 |
0 |
75 |
| PpuM I |
4 |
0 |
25 |
0 |
100 |
| PshA I |
4 + BSA |
50 |
75 |
10 |
100 |
| Psi I * |
SE-B |
50 |
100 |
0 |
25 |
| PspG I @75°C |
2 |
75 |
100 |
50 |
100 |
| PspOM I * |
SE-Y |
25 |
100 |
50 |
100 |
| Pst I |
3 + BSA |
75 |
75 |
100 |
50 |
| Pvu I |
3 + BSA |
10 |
75 |
100 |
10 |
| Pvu II |
2 |
100 |
100 |
100 |
100 |
| Rsa I |
1 |
100 |
100 |
50 |
100 |
| Rsr II |
4 |
25 |
75 |
10 |
100 |
| Sac I |
1 + BSA |
100 |
50 |
10 |
100 |
| Sac II |
4 |
25 |
75 |
10 |
100 |
| Sal I |
U + BSA |
0 |
0 |
50 |
0 |
| Sap I |
4 |
75 |
50 |
0 |
100 |
| Sau3A I |
U + BSA |
100 |
50 |
10 |
100 |
| Sau96 I |
4 |
50 |
100 |
100 |
100 |
| Sbf I * |
SE-Y |
75 |
50 |
0 |
100 |
| Sca I |
U |
25 |
75 |
dd50 |
25 |
| ScrF I |
4 |
100 |
100 |
100 |
100 |
| SexA I |
3 + BSA |
25 |
25 |
100 |
100 |
| SfaN I |
3 |
0 |
75 |
100 |
25 |
| Sfc I |
4 + BSA |
75 |
50 |
10 |
100 |
| Sfi I @50°C |
2 + BSA |
0 |
100 |
10 |
100 |
| Sfo I |
2 |
25 |
100 |
50 |
50 |
| SgrA I |
4 |
100 |
50 |
10 |
100 |
| |