Question

The very useful restriction enzyme SpoBob hydrolyses the phosphodiester bond between the guanylates in the sequence AGGACCA. Is SpoBob generating blunt or sticky ends? Please explain.

Answer 1

Ans.

The restriction enzyme will cut at the recognition sequence, regardless of the number of times it appears.
Some restriction enzymes can cut a similar but not exact sequence if the incubation with the enzyme is prolonged and or if the enzyme is present at a high concentration.
Eg: Restriction fragment length polymorphism (RFLP) is a method that analyzes banding patterns from DNA cut by restriction endonucleases (RE). DNA is incubated with appropriate restriction enzyme and then separated by electrophoresis. The restriction enzyme will cut all sequences which had to be identify and then we will have several DNA bands. These RE sequences will usually appear at different areas of different individual genomes and will get different bands between samples.
Some Bacterial strains and other organisms can also be "typed" or characterized similarly because restriction endonucleases (RE) will not cut at mutated RE recognition sequences hence different restriction banding patterns.
In the following sequence, how many double-stranded cuts will take place, and will the cuts result in blunt or sticky ends? TCGAAAGCTTCGTATCGTAAGCTTGCT.