内切酶的热失活参数
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热失活是终止酶切反应的一种简便易行的方法。大多数最适反应温度是37℃的酶在65℃下温育20分钟即可失活。一些在65℃下不能失活的酶如将温度升高至80℃，温育20分钟也可失活。下表注明了每种酶的失活条件。
在50μl的反应体系中，37℃条件下，以0.5μg小牛胸腺DNA作为底物，加入5-10μl内切酶及相应缓冲液，反应60分钟，然后升温至65℃或80℃温育20分钟进行热失活。在上述混合液中加入0.5μg底物DNA（通常为λDNA），调节温度至最适反应温度，温育60分钟。若琼脂糖电泳显示底物部分或完全被消化，则表明该酶没有完全被热失活。
| Enzyme | Heat Inactivation (temperature) |
| Aat II | Yes (65°C) |
| Acc I | Yes (80°C) |
| Acc65 I | Yes (65°C) |
| Aci I | Yes (65°C) |
| Acl I | No |
| Acu I | Yes (65°C) |
| Afe I | Yes (65°C) |
| Afl II | Yes (65°C) |
| Afl III | Yes (80°C) |
| Age I | Yes (65°C) |
| Ahd I | Yes (65°C) |
| Ale I | Yes (65°C) |
| Alu I | Yes (65°C) |
| Alw I | Yes (65°C) |
| AlwN I | Yes (65°C) |
| Apa I | Yes (65°C) |
| ApaL I | No |
| ApeK I | No |
| Apo I | Yes (80°C) |
| Asc I | Yes (65°C) |
| Ase I | Yes (65°C) |
| AsiS I | Yes (80°C) |
| Ava I | Yes (80°C) |
| Ava II | Yes (65°C) |
| Avr II | No |
| Bae I | Yes (65°C) |
| BamH I | No |
| Ban I | Yes (65°C) |
| Ban II | Yes (65°C) |
| Bbs I | Yes (65°C) |
| Bbv I | Yes (65°C) |
| BbvC I | Yes (80°C) |
| Bcc I | Yes (65°C) |
| BceA I | Yes (65°C) |
| Bcg I | Yes (65°C) |
| BciV I | Yes (65°C) |
| Bcl I | No |
| Bfa I | Yes (80°C) |
| BfrB I | Yes (80°C) |
| BfuA I | Yes (65°C) |
| BfuC I | Yes (80°C) |
| Bgl I | Yes (65°C) |
| Bgl II | No |
| Blp I | No |
| Bme1580 I | Yes (80°C) |
| BmgB I | Yes (65°C) |
| Bmr I | Yes (65°C) |
| Bmt I | Yes (65°C) |
| Bpm I | Yes (65°C) |
| Bpu10 I | Yes (80°C) |
| BpuE I | Yes (65°C) |
| Bsa I | Yes (65°C) |
| BsaA I | Yes (80°C) |
| BsaB I | Yes (80°C) |
| BsaH I | Yes (80°C) |
| BsaJ I | Yes (80°C) |
| BsaW I | Yes (80°C) |
| BsaX I | No |
| BseR I | Yes (65°C) |
| BseY I | Yes (65°C) |
| Bsg I | Yes (65°C) |
| BsiE I | Yes (80°C) |
| BsiHKA I | Yes (80°C) |
| BsiW I | Yes (80°C) |
| Bsl I | Yes (80°C) |
| Bsm I | Yes (80°C) |
| BsmA I | Yes (80°C) |
| BsmB I | Yes (80°C) |
| BsmF I | Yes (80°C) |
| BsoB I | No |
| Bsp1286 I | Yes (65°C) |
| BspCN I | Yes (80°C) |
| BspD I | Yes (65°C) |
| BspE I | Yes (80°C) |
| BspH I | Yes (65°C) |
| BspM I | Yes (65°C) |
| Bsr I | Yes (80°C) |
| BsrB I | Yes (80°C) |
| BsrD I | Yes (80°C) |
| BsrF I | No |
| BsrG I | Yes (80°C) |
| BssH II | Yes (80°C) |
| BssK I | Yes (80°C) |
| BssS I | Yes (80°C) |
| BstAP I | Yes (80°C) |
| BstB I | No |
| BstE II | No |
| BstF5 I | No |
| BstN I | No |
| BstU I | No |
| BstX I | Yes (65°C) |
| BstY I | Yes (80°C) |
| BstZ17 I | No |
| Bsu36 I | Yes (80°C) |
| Btg I | Yes (80°C) |
| BtgZ I | Yes (80°C) |
| Bts I | Yes (80°C) |
| Cac8 I | Yes (65°C) |
| Cla I | Yes (65°C) |
| CviA II | Yes (65°C) |
| Dde I | Yes (65°C) |
| Dpn I | Yes (80°C) |
| Dpn II | Yes (65°C) |
| Dra I | Yes (65°C) |
| Dra III | Yes (65°C) |
| Drd I | Yes (65°C) |
| Eae I | Yes (65°C) |
| Eag I | Yes (65°C) |
| Ear I | Yes (65°C) |
| Eci I | Yes (65°C) |
| EcoN I | Yes (65°C) |
| EcoO109 I | Yes (65°C) |
| EcoR I | Yes (65°C) |
| EcoR V | Yes (80°C) |
| Fat I | Yes (65°C) |
| Fau I | Yes (65°C) |
| Fnu4H I | Yes (65°C) |
| Fok I | Yes (65°C) |
| Fse I | Yes (65°C) |
| Fsp I | Yes (65°C) |
| Hae II | Yes (80°C) |
| Hae III | Yes (80°C) |
| Hga I | Yes (65°C) |
| Hha I | Yes (65°C) |
| HinP1 I | Yes (65°C) |
| Hinc II | Yes (65°C) |
| Hind III | Yes (65°C) |
| Hinf I | Yes (80°C) |
| Hpa I | No |
| Hpa II | Yes (65°C) |
| Hph I | Yes (65°C) |
| Hpy99 I | Yes (65°C) |
| Hpy188 I | Yes (65°C) |
| Hpy188 III | Yes (65°C) |
| HpyCH4 III | Yes (80°C) |
| HpyCH4 IV | Yes (65°C) |
| HpyCH4 V | Yes (65°C) |
| I-Ceu I | Yes (65°C) |
| I-Sce I | Yes (65°C) |
| Kas I | Yes (65°C) |
| Kpn I | No |
| Mbo I | Yes (65°C) |
| Mbo II | Yes (65°C) |
| McrBC | Yes (65°C) |
| Mfe I | Yes (65°C) |
| Mlu I | Yes (65°C) |
| Mly I | Yes (65°C) |
| Mme I | Yes (80°C) |
| Mnl I | Yes (65°C) |
| Msc I | Yes (65°C) |
| Mse I | Yes (65°C) |
| Msl I | Yes (65°C) |
| Msp I | Yes (65°C) |
| MspA1 I | Yes (65°C) |
| Mwo I | No |
| Nae I | Yes (65°C) |
| N.Alw I | Yes (80°C) |
| Nar I | Yes (65°C) |
| N.BbvC IA | Yes (80°C) |
| N.BbvC IB | Yes (80°C) |
| N.BstNB I | Yes (80°C) |
| Nci I | No |
| Nco I | Yes (65°C) |
| Nde I | Yes (65°C) |
| NgoM IV | Yes (80°C) |
| Nhe I | Yes (65°C) |
| Nla III | Yes (65°C) |
| Nla IV | Yes (65°C) |
| Not I | Yes (65°C) |
| Nru I | Yes (65°C) |
| Nsi I | Yes (65°C) |
| Nsp I | Yes (65°C) |
| Pac I | Yes (65°C) |
| PaeR7 I | No |
| Pci I | Yes (65°C) |
| PflF I | Yes (65°C) |
| PflM I | Yes (65°C) |
| Pho I | No |
| PI-Psp I | No |
| Pl-Sce I | Yes (65°C) |
| Ple I | Yes (65°C) |
| Pme I | Yes (65°C) |
| Pml I | Yes (65°C) |
| PpuM I | No |
| PshA I | Yes (65°C) |
| Psi I | Yes (65°C) |
| PspG I | No |
| PspOM I | Yes (65°C) |
| Pst I | Yes (80°C) |
| Pvu I | Yes (80°C) |
| Pvu II | No |
| Rsa I | Yes (65°C) |
| Rsr II | Yes (65°C) |
| Sac I | Yes (65°C) |
| Sac II | Yes (65°C) |
| Sal I | Yes (65°C) |
| Sap I | Yes (65°C) |
| Sau96 I | Yes (80°C) |
| Sau3A I | Yes (65°C) |
| Sbf I | Yes (65°C) |
| Sca I | Yes (80°C) |
| ScrF I | Yes (65°C) |
| SexA I | Yes (65°C) |
| SfaN I | Yes (65°C) |
| Sfc I | Yes (65°C) |
| Sfi I | No |
| Sfo I | Yes (65°C) |
| SgrA I | Yes (65°C) |
| Sma I | Yes (65°C) |
| Sml I | No |
| SnaB I | Yes (80°C) |
| Spe I | Yes (65°C) |
| Sph I | Yes (65°C) |
| Ssp I | Yes (65°C) |
| Stu I | Yes (65°C) |
| Sty I | Yes (65°C) |
| StyD4 I | Yes (65°C) |
| Swa I | Yes (65°C) |
| Taq I | Yes (80°C) |
| Tfi I | No |
| Tli I | No |
| Tse I | No |
| Tsp45 I | No |
| Tsp509 I | No |
| TspR I | No |
| Tth111 I | No |
| Xba I | Yes (65°C) |
| Xcm I | Yes (65°C) |
| Xho I | Yes (65°C) |
| Xma I | Yes (65°C) |
| Xmn I | Yes (65°C) |
| Zra I | Yes (80°C) |
