3.4 Heat-Inactivation of Restriction Enzymes
Heat-inactivation of restriction enzymes may be performed when a subsequent reaction
can be performed in the same reaction buffer, or when the reaction will be diluted for the
next application. This will eliminate the need for extra ethanol precipitations or
clean-up steps. Table 3.4 lists the sensitivity of Promega's restriction enzymes to
heat-inactivation.
Promega
Enzyme |
Heat
Inactivated |
| Aat II |
+ |
| Acc I |
- |
| Acc III |
- |
| Acc65 I |
+ |
| AccB7 I |
+ |
| Age I |
+ |
| Alu I |
+ |
| Alw26 I |
+ |
| Alw44 I |
+ |
| Apa I |
+ |
| Ava I |
+/- |
| Ava II |
+ |
| Bal I |
+ |
| BamH I |
+ |
| Ban I |
- |
| Ban II |
+ |
| Bbu I |
+ |
| Bcl I |
- |
| Bgl I |
+ |
| Bgl II |
- |
| BsaM I |
- |
| BsaO I |
- |
| Bsp1286 I |
+ |
| BsrBR I |
- |
| BsrS I |
- |
| BssH II |
- |
| Bst71 I |
- |
| Bst98 I |
- |
| BstE II |
- |
| BstO I |
- |
| BstX I |
+/- |
| BstZ I |
- |
| Bsu36 I |
- |
| Cfo I |
+/- |
| Cla I |
+ |
| Csp I |
+ |
| Csp45 I |
+ |
| Dde I |
+/- |
| Dpn I |
+ |
| Dra I |
+ |
| EclHK I |
+ |
| Eco47 III |
+ |
| Eco52 I |
+ |
| EcoICR I |
+ |
| EcoR I |
+ |
| EcoR V |
+ |
| Fok I |
+ |
| Hae II |
- |
| Hae III |
- |
| Hha I |
+ |
| Hinc II |
+ |
| Hind III |
+ |
|
|
Promega
Enzyme |
Heat
Inactivated |
| Hinf I |
- |
| Hpa I |
- |
| Hpa II |
- |
| Hsp92 I |
+ |
| Hsp92 II |
+ |
| I-Ppo I |
+ |
| Kpn I |
+/- |
| Mbo I |
+ |
| Mbo II |
+ |
| Mlu I |
+/- |
| Msp I |
+ |
| MspA1 I |
+ |
| Nae I |
+ |
| Nar I |
+ |
| Nci I |
+ |
| Nco I |
+ |
| Nde I |
+ |
| Nde II |
+ |
| NgoM IV |
+ |
| Nhe I |
+ |
| Not I |
+ |
| Nru I |
+ |
| Nsi I |
+/- |
| Pst I |
+ |
| Pvu I |
- |
| Pvu II |
+ |
| Rsa I |
+ |
| Sac I |
+ |
| Sac II |
+ |
| Sal I |
+ |
| Sau3A I |
+ |
| Sau96 I |
- |
| Sca I |
+ |
| Sfi I |
- |
| Sgf I(a) |
+/- |
| Sin I |
+ |
| Sma I |
+ |
| SnaB I |
- |
| Spe I |
+ |
| Sph I |
+ |
| Ssp I |
+ |
| Stu I |
+ |
| Sty I |
+ |
| Taq I |
- |
| Tru9 I |
- |
| Tth111 I |
- |
| Vsp I |
+ |
| Xba I |
- |
| Xho I |
+ |
| Xho II |
+ |
| Xma I |
+ |
| Xmn I |
+ |
|
Key:
| + |
greater than 95% inactivation
(DNA is undigested). |
| - |
less that 95% inactivation (DNA digest is complete,
i.e., at least 5% of the initial 20 activity units [at least 1 unit] remain(s).) |
| +/- |
partial inactivation (DNA is partially digested). |
Conditions: Twenty units of enzyme in 50µl of its optimal buffer were heated at 65°C
for 15 minutes. 1µg of DNA was added and incubated for 1 hour in accordance with the unit
definition, then analyzed by agarose gel electrophoresis.
|
|
