| 各种失活处理后限制酶的残存活性 |
| |
|
|
限制酶酶切反应后,通常使用热处理的方式使限制酶失活。但是,由于不同的限制酶耐热性不同,并且考虑到DNA的变性温度,仅通过热处理的方式是不足以将限制酶完全失活的。
因此,我们对每种酶进行了四种失活处理并检测其残存活性,以检讨完全失活的条件(该数据不适用于QuickCut系列限制酶)。 |
| |
| 1.60℃加热处理15 min; 2. 70℃加热处理15 min; 3.乙醇沉淀处理; 4.酚处理-乙醇沉淀处理 |
| |
| +:有残存活性 -:无残存活性 NT:未检测 |
| |
| 限制酶 |
60℃, 15 min |
70℃, 15 min |
乙醇沉淀处理 |
酚处理-乙醇沉淀处理 |
| Aat II |
+
|
-
|
-
|
NT
|
| Acc I |
+
|
-
|
-
|
NT
|
| Acc II |
+
|
+
|
-
|
NT
|
| Acc III |
NT
|
-
|
-
|
NT
|
| Afa I |
+
|
-
|
+
|
-
|
| Afl II |
-
|
-
|
-
|
-
|
| Alu I |
-
|
NT
|
+
|
-
|
| Aor13H I |
NT
|
+
|
+
|
-
|
| Aor51H I |
-
|
NT
|
-
|
NT
|
| Apa I |
-
|
NT
|
-
|
NT
|
| ApaL I |
+
|
-
|
-
|
NT
|
| Bal I |
-
|
-
|
-
|
-
|
| BamH I |
+
|
+
|
-
|
-
|
| Ban II |
-
|
NT
|
-
|
NT
|
| Bci T130 I |
+
|
+
|
-
|
-
|
| Bcn I |
+
|
+
|
-
|
NT
|
| Bgl I |
-
|
NT
|
+
|
-
|
| Bgl II |
+
|
-
|
+
|
-
|
| Bln I |
+
|
+
|
-
|
-
|
| BmeT110 I |
-
|
NT
|
-
|
NT
|
| BmgT120 I |
+
|
+
|
-
|
NT
|
| Bpu1102 I |
+
|
+
|
+
|
-
|
| BspT104 I |
+
|
+
|
-
|
-
|
| BspT107 I |
-
|
NT
|
-
|
NT
|
| Bsp1286 I |
-
|
NT
|
-
|
NT
|
| Bsp1407 I |
+
|
-
|
+
|
-
|
| BssH II |
+
|
-
|
-
|
NT
|
| BstP I |
NT
|
+
|
-
|
NT
|
| BstX I |
-
|
NT
|
-
|
NT
|
| Bst1107 I |
+
|
+
|
+
|
-
|
| Cfr10 I |
+
|
+
|
+
|
-
|
| Cla I |
+
|
-
|
-
|
-
|
| Cpo I |
-
|
NT
|
-
|
NT
|
| Dde I |
+
|
-
|
-
|
-
|
| Dra I |
-
|
NT
|
-
|
NT
|
| Dpn I |
+
|
-
|
+
|
-
|
| Eae I |
-
|
NT
|
-
|
NT
|
| Eam1105 I |
-
|
NT
|
+
|
-
|
| EcoO65 I |
+
|
-
|
+
|
-
|
| EcoO109 I |
-
|
NT
|
-
|
NT
|
| EcoR I |
-
|
NT
|
-
|
NT
|
| EcoR V |
+
|
-
|
+
|
-
|
| EcoT14 I |
-
|
NT
|
-
|
NT
|
| EcoT22 I |
-
|
NT
|
-
|
NT
|
| Eco52 I |
-
|
NT
|
+
|
-
|
| Eco81 I |
+
|
-
|
-
|
NT
|
| Fba I |
+
|
+
|
-
|
NT
|
| Fok I |
-
|
NT
|
+
|
-
|
| Hae II |
-
|
-
|
-
|
-
|
| Hae III |
+
|
-
|
-
|
NT
|
| Hap II |
+
|
-
|
+
|
-
|
|
| |
| 限制酶 |
60℃, 15 min |
70℃, 15 min |
乙醇沉淀处理 |
酚处理-乙醇沉淀处理 |
| Hha I |
+
|
-
|
-
|
-
|
| Hinc II |
+
|
-
|
-
|
NT
|
| Hind III |
+
|
-
|
-
|
NT
|
| Hinf I |
+
|
-
|
-
|
NT
|
| Hin1 I |
-
|
NT
|
+
|
-
|
| Hpa I |
-
|
NT
|
-
|
NT
|
| Kpn I |
-
|
NT
|
-
|
NT
|
| Mbo I |
+
|
-
|
-
|
-
|
| Mbo II |
+
|
-
|
-
|
NT
|
| Mfl I |
+
|
+
|
+
|
-
|
| Mlu I |
+
|
+
|
-
|
NT
|
| Msp I |
+
|
+
|
+
|
-
|
| Mun I |
+
|
-
|
+
|
-
|
| Nae I |
-
|
NT
|
-
|
NT
|
| Nco I |
+
|
-
|
+
|
-
|
| Nde I |
-
|
-
|
-
|
-
|
| Nhe I |
+
|
-
|
-
|
NT
|
| Not I |
-
|
-
|
+
|
-
|
| Nru I |
-
|
NT
|
+
|
-
|
| Nsb I |
+
|
-
|
+
|
-
|
| PmaC I |
-
|
NT
|
-
|
NT
|
| PshA I |
-
|
NT
|
-
|
NT
|
| PshB I |
+
|
-
|
-
|
NT
|
| Psp1406 I |
-
|
NT
|
+
|
-
|
| Pst I |
-
|
NT
|
-
|
NT
|
| Pvu I |
-
|
-
|
-
|
-
|
| Pvu II |
+
|
+
|
+
|
-
|
| RspRS II |
NT
|
+
|
+
|
-
|
| Sac I |
-
|
NT
|
-
|
NT
|
| Sac II |
+
|
-
|
+
|
-
|
| Sal I |
-
|
-
|
-
|
-
|
| Sau3A I |
+
|
-
|
-
|
NT
|
| Sca I |
-
|
NT
|
-
|
NT
|
| Sfi I |
+
|
-
|
+
|
-
|
| Sma I |
-
|
NT
|
+
|
-
|
| Smi I |
+
|
+
|
+
|
-
|
| SnaB I |
-
|
NT
|
-
|
NT
|
| Spe I |
-
|
NT
|
-
|
NT
|
| Sph I |
-
|
NT
|
-
|
NT
|
| Sse8387 I |
-
|
NT
|
-
|
NT
|
| Ssp I |
-
|
NT
|
-
|
NT
|
| Stu I |
-
|
NT
|
+
|
-
|
| Taq I |
NT
|
+
|
+
|
-
|
| Tth111 I |
NT
|
+
|
-
|
NT
|
| Van91 I |
-
|
NT
|
+
|
-
|
| VpaK11B I |
-
|
NT
|
-
|
NT
|
| Xba I |
-
|
-
|
-
|
-
|
| Xho I |
+
|
+
|
-
|
-
|
| Xsp I |
+
|
+
|
+
|
-
|
|
| |
| |
| |
| |
|