Final answer:
EcoRI, a type II restriction enzyme, typically requires about 1 hour to digest DNA at 37°C under optimal conditions. The time can vary depending on variables such as DNA and enzyme concentrations. EcoRI is widely used in recombinant DNA technology, cutting at a specific recognition site to enable DNA fragment manipulation.
Step-by-step explanation:
EcoRI is a type II restriction enzyme derived from the bacterium Escherichia coli, which recognizes the specific DNA sequence 5'GAATTC3' and cuts at this site to produce 'sticky ends'. The optimal conditions for EcoRI activity generally include a specific temperature, usually 37°C, and a buffer that provides necessary ions and optimal pH. According to most protocols, such as those from New England Biolabs, restriction endonuclease digests, including those using EcoRI, typically take about 1 hour. However, the time can vary depending on the concentration of DNA, the enzyme, buffer conditions, and the amount of DNA to be cut.
It's important to set up reactions correctly and ensure that enzymes are kept ice-cold before use to maintain activity. A typical use for EcoRI and other enzymes involves cutting plasmid DNA for recombinant DNA technology applications, where DNA fragments can then be ligated into vectors for cloning or other genetic manipulations. After the digestion, the reactions can be stored at -20°C if not immediately used.