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AAT Bioquest

What are the factors that affect the activity of restriction enzymes?

Posted July 6, 2023


Answer

The activity of restriction enzymes are affected by the following factors: 

Temperature - Most restriction enzymes work most efficiently at 37°C, but there are a few exceptions. Some work best at higher temperatures ranging from 50°C to 65°C, while a few cut best at lower temperatures of about 25°C.

Buffer systems - Most restriction enzymes exhibit maximum activity in the pH range of 7.0–8.0. 

Ionic Conditions - Most restriction enzymes need Mg2+ but a few also require ions such as Na+ and K+.

Cofactors - Most restriction enzymes need certain cofactors or combinations of cofactors to facilitate digestion at the recognition site. All enzymes require Mg2+ as a cofactor for the endonuclease activity. 

Methylation status of DNA - DNA digestion may be affected by the methylation of adenine or cytidine residues.

Additional resources

Some factors affecting the action of restriction endonucleases on human metaphase chromosomes

Restriction Enzymes Cut Sites Reference Table

Enzymes

6-ROX glycine *25 uM fluorescence reference solution for PCR reactions*