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长时间反应时内切酶的活性保持情况因酶而异。本表中列出了在16小时内完全酶解底物DNA所需的最小酶量。
实验方法：在50µl的反应体系中，分别加入1µg的单位定义底物DNA和1.00、0.50、0.25和0.13个单位的内切酶，37°C（或要求的最适温度）反应16小时。用琼脂糖凝胶电泳法来确定在16小时内酶解1µg底物DNA所需的最小酶量。
16小时酶解1µg底物需要酶量小于1单位的酶可以用加入较少的酶量和延长反应时间的方法节省酶的用量。实际16小时酶解1µg底物的内切酶最小用量视底物的不同而有所变化。
| 酶 |
16小时反应所需的最小酶量 |
| Aat II |
0.13 |
| Acc I |
0.13 |
| Acc65 I |
0.50 |
| Aci I |
1.00 |
| Acl I |
0.50 |
| Acu I |
1.00 |
| Afe I |
0.25 |
| Afl II |
0.13 |
| Afl III |
0.13 |
| Age I |
1.00 |
| Ahd I |
0.13 |
| Ale I |
0.25 |
| Alu I |
0.25 |
| Alw I |
0.50 |
| AlwN I |
0.13 |
| Apa I @25°C |
0.13 |
| ApaL I |
1.00 |
| ApeK I @75°C |
0.13 |
| Apo I @50°C |
0.13 |
| Asc I |
0.13 |
| Ase I |
0.13 |
| AsiS I |
0.13 |
| Ava I |
0.25 |
| Ava II |
0.25 |
| Avr II |
0.13 |
| Bae I@25°C |
0.50 |
| BamH I |
0.50 |
| Ban I |
0.13 |
| Ban II |
0.50 |
| Bbs I |
0.25 |
| Bbv I |
1.00 |
| BbvC I |
0.13 |
| Bcc I |
0.50 |
| BceA I |
0.13 |
| Bcg I |
0.50 |
| BciV I |
1.00 |
| Bcl I @50°C |
0.50 |
| Bfa I |
0.50 |
| BfrB I |
0.50 |
| BfuA I @50°C |
0.25 |
| BfuC I |
1.00 |
| Bgl I |
0.13 |
| Bgl II |
0.25 |
| Blp I |
0.50 |
| Bme1580 I |
0.50 |
| BmgB I |
1.00 |
| Bmr I |
1.00 |
| Bmt I |
n/a |
| Bpm I |
1.00 |
| Bpu10 I |
0.50 |
| BpuE I |
1.00 |
| Bsa I @50°C |
1.00 |
| BsaA I |
1.00 |
| BsaB I @60°C |
0.13 |
| BsaH I |
0.13 |
| BsaJ I @60°C |
0.13 |
| BsaW I @60°C |
0.13 |
| BsaX I |
0.25 |
| BseR I |
1.00 |
| BseY I |
0.25 |
| Bsg I |
0.50 |
| BsiE I @60°C |
0.25 |
| BsiHKA I @65°C |
1.00 |
| BsiW I @55°C |
0.50 |
| Bsl I @55°C |
0.13 |
| Bsm I @65°C |
0.50 |
| BsmA I @55°C |
0.25 |
| BsmB I @55°C |
0.50 |
| BsmF I @65°C |
0.13 |
| BsoB I |
0.13 |
| Bsp1286 I |
0.50 |
| BspCN I @25°C |
1.00 |
| BspD I |
0.25 |
| BspE I |
0.13 |
| BspH I |
0.13 |
| BspM I |
0.25 |
| Bsr I @65°C |
0.25 |
| BsrB I |
0.13 |
| BsrD I @65°C |
0.50 |
| BsrF I |
0.13 |
| BsrG I |
0.13 |
| BssH II @50°C |
0.50 |
| BssK I @60°C |
0.25 |
| BssS I |
0.25 |
| BstAP I @60°C |
0.50 |
| BstB I @65°C |
0.13 |
| BstE II @60°C |
1.00 |
| BstF5 I @65°C |
0.25 |
| BstN I @60°C |
1.00 |
| BstU I @60°C |
0.13 |
| BstX I @55°C |
0.25 |
| BstY I @60°C |
0.13 |
| BstZ17 I |
1.00 |
| Bsu36 I |
0.13 |
| Btg I |
0.50 |
| BtgZ I |
1.00 |
| Bts I |
0.50 |
| Cac8 I |
0.25 |
| Cla I |
0.50 |
| CviA II |
1.00 |
| Dde I |
0.13 |
| Dpn I |
0.13 |
| Dpn II |
0.13 |
| Dra I |
0.25 |
| Dra III |
0.25 |
| Drd I |
0.13 |
| Eae I |
0.13 |
| Eag I |
0.50 |
| Ear I |
0.13 |
| Eci I |
1.00 |
| EcoN I |
0.13 |
| EcoO109 I |
0.13 |
| EcoR I |
0.13 |
| EcoR V |
0.50 |
| Fat I |
n/a |
| Fau I @55°C |
0.50 |
| Fnu4H I |
0.25 |
| Fok I |
1.00 |
| Fse I |
1.00 |
| Fsp I |
0.13 |
| Hae II |
1.00 |
| Hae III |
0.13 |
| Hga I |
1.00 |
| Hha I |
0.25 |
| Hinc II |
0.13 |
| Hind III |
0.13 |
| Hinf I |
0.13 |
| HinP1 I |
0.13 |
| Hpa I |
0.25 |
| Hpa II |
0.25 |
| Hph I |
0.25 |
| Hpy99 I |
1.00 |
| Hpy188 I |
1.00 |
| Hpy188 III |
0.13 |
| HpyCH4 III |
0.13 |
| HpyCH4 IV |
0.25 |
| HpyCH4 V |
0.25 |
| I-Ceu I |
0.25 |
| I-Sce I |
0.25 |
| Kas I |
1.00 |
| Kpn I |
0.25 |
| Mbo I |
0.50 |
| Mbo II |
1.00 |
| McrBC |
1.00 |
| Mfe I |
0.25 |
| Mlu I |
0.13 |
| Mly I |
1.00 |
| Mme I |
1.00 |
| Mnl I |
0.25 |
| Msc I |
0.50 |
| Mse I |
0.13 |
| Msl I |
1.00 |
| Msp I |
0.50 |
| MspA1 I |
0.25 |
| Mwo I @60°C |
0.13 |
| Nae I |
0.50 |
| N.Awl I |
0.13 |
| Nar I |
1.00 |
| N.BbvC IA |
0.13 |
| N.BbvC IB |
0.13 |
| Nci I |
0.50 |
| Nco I |
0.25 |
| Nde I |
0.13 |
| NgoM IV |
0.13 |
| Nhe I |
0.25 |
| Nla III |
0.50 |
| Nla IV |
0.50 |
| Not I |
0.25 |
| Nru I |
0.13 |
| Nsi I |
0.50 |
| Nsp I |
0.25 |
| Pac I |
0.13 |
| PaeR7 I |
0.13 |
| Pci I |
0.25 |
| PflF I |
0.13 |
| PflM I |
1.00 |
| Pho I |
0.13 |
| PI-Pso I @65°C |
0.13 |
| PI-Sce I |
0.13 |
| Ple I |
1.00 |
| Pme I |
1.00 |
| Pml I |
0.50 |
| PpuM I |
0.13 |
| PshA I |
1.00 |
| Psi I |
0.25 |
| PspG I @75°C |
0.25 |
| PspOM I |
0.13 |
| Pst I |
0.50 |
| Pvu I |
1.00 |
| Pvu II |
0.13 |
| Rsa I |
0.13 |
| Rsr II |
0.25 |
| Sac I |
0.13 |
| Sac II |
0.13 |
| Sal I |
0.13 |
| Sap I |
1.00 |
| Sau3A I |
0.50 |
| Sau96 I |
0.25 |
| Sbf I |
0.50 |
| Sca I |
0.25 |
| ScrF I |
0.25 |
| SexA I |
0.25 |
| SfaN I |
0.50 |
| Sfc I |
1.00 |
| Sfi I @50°C |
0.25 |
| Sfo I |
0.50 |
| SgrA I |
1.00 |
| Sma I @25°C |
0.13 |
| Sml I @55°C |
0.25 |
| SnaB I |
0.50 |
| Spe I |
0.50 |
| Sph I |
0.13 |
| Ssp I |
0.25 |
| Stu I |
0.25 |
| Sty I |
0.25 |
| StyD4 I |
0.13 |
| Swa I @25°C |
0.25 |
| Taq I @65°C |
0.50 |
| Tfi I @65°C |
0.25 |
| Tli I @75°C |
0.25 |
| Tse I @65°C |
0.25 |
| Tsp45 I @65°C |
0.50 |
| Tsp509 I @65°C |
0.13 |
| TspR I @65°C |
0.13 |
| Tth111 I @65°C |
0.25 |
| Xba I |
0.13 |
| Xcm I |
0.13 |
| Xho I |
0.13 |
| Xma I |
0.50 |
| Xmn I |
0.25 |
| Zra I |
n/a |
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