各種失活處理后限制酶的殘存活性 |
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限制酶酶切反應(yīng)后,通常使用熱處理的方式使限制酶失活。但是,由于不同的限制酶耐熱性不同,并且考慮到DNA的變性溫度,僅通過熱處理的方式是不足以將限制酶完全失活的。
因此,我們對每種酶進(jìn)行了四種失活處理并檢測其殘存活性,以檢討完全失活的條件(該數(shù)據(jù)不適用于QuickCut系列限制酶)。 |
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1.60℃加熱處理15 min; 2. 70℃加熱處理15 min; 3.乙醇沉淀處理; 4.酚處理-乙醇沉淀處理 |
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+:有殘存活性 -:無殘存活性 NT:未檢測 |
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限制酶 |
60℃, 15 min |
70℃, 15 min |
乙醇沉淀處理 |
酚處理-乙醇沉淀處理 |
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
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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
|
+
|
-
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Eco81 I |
+
|
-
|
-
|
NT
|
Fba I |
+
|
+
|
-
|
NT
|
Fok I |
-
|
NT
|
+
|
-
|
Hae II |
-
|
-
|
-
|
-
|
Hae III |
+
|
-
|
-
|
NT
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Hap II |
+
|
-
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+
|
-
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限制酶 |
60℃, 15 min |
70℃, 15 min |
乙醇沉淀處理 |
酚處理-乙醇沉淀處理 |
Hha I |
+
|
-
|
-
|
-
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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 |
+
|
+
|
+
|
-
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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 |
+
|
+
|
+
|
-
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|
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