Restriction Fragment Length Polymorphism (RFLP)

Question 1

RFLP

RFLP analysis?

Answer 1

• Compares different lengths of DNA fragments made by restriction endonucleouses
• large quantities of DNA will smear after gel electrophoresis

Question 2

RFLP

To detect differences? 4 steps?

Answer 2

• placed on nylon membrane and chemical added to denture DNA into single strands
• electric current used to cause single strand DNA to migrate onto nylon membrane (southern blotting)
• nylon membrane soaked in solution containing complimentary radioactive probes designed to anneal to regions in process called hybrodization
• nylon then placed against X-ray film, autoradiogram produced

Question 3

RFLP

Polymorphism?

Answer 3

Any difference in DNA that can be detected between individuals can be present in both coding and non coding regions