Heat Inactivation of Restriction Enzymes
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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 cleanup steps. This reference table lists the sensitivity of Promega's restriction enzymes to heat-inactivation.
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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] remains).
+/– 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. One microgram of DNA was added and incubated for 1 hour in accordance with the unit
definition and then analyzed by agarose gel electrophoresis.