Heat Inactivation of Restriction Enzymes

Equipment & Consumables:

• Heat Block

• Your Restriction Enzyme Digest solutions

• A web resource that explains the inactivation temperature of your specific enzyme brand, such as this one.

Protocol:

1. Determine whether your enzymes can be heat inactivated by visiting the website of your enzyme provider. In my case, this is NEB’s website.

   • Some brands produce slightly different versions of each enzyme that may be more/less susceptible to heat inactivation.

2. If your enzymes can both be inactivated, set your heat block to the higher temperature and place your sample in the heat block for 20 minutes.

3. Proceed to the Ligation protocol to stick your fragments together, or Agarose Electrophoresis for immediate analysis (or further purification if Heat Inactivation was insufficient).

Acknowledgements:

• Coleman Protocols 2017 + 2019 http://coleman-lab.org/

• https://www.nebiolabs.com.au/tools-and-resources/usage-guidelines/heat-inactivation