Final answer:
EcoRI is a Type II restriction enzyme that cleaves DNA at a specific recognition sequence, typically requiring approximately one hour to digest DNA under optimal conditions. These conditions often involve a temperature of 37 degrees Celsius and a specific reaction buffer.
Step-by-step explanation:
Restriction enzymes are integral tools in molecular biology, particularly in recombinant DNA technology. EcoRI is a well-known Type II restriction enzyme derived from the bacterium Escherichia coli. This enzyme recognizes the specific DNA sequence 5'GA-A-T-T-C3' and cleaves the DNA between the G and A nucleotides to create 'sticky ends'. These sticky ends can pair with complementary DNA sequences of a vector during the cloning process, where a ligase enzyme will subsequently 'glue' the annealed fragments together.
The action of EcoRI and its cleavage specificity allow for the creation of a DNA fingerprint that is unique to each DNA sample, making it an essential tool for genetic analysis. In laboratory settings, whether in educational settings such as a high school lab or in research institutions, the typical protocol suggests that EcoRI and other restriction endonucleases digest DNA in approximately one hour under optimal conditions. This duration allows sufficient time for EcoRI to scan the DNA, find its specific recognition site, and make the cut. The conditions for EcoRI activity typically include a temperature of 37℃ and a specific buffer provided by enzyme suppliers, which provides the necessary ions and pH for enzyme activity.
It's important to note, though, that the specific digestion time can depend on various factors including the concentration of the enzyme, the purity and amount of DNA substrate, and the precise temperature of the reaction. Generally, reliable suppliers like New England Biolabs offer detailed guidelines for the use of EcoRI and other restriction enzymes.