Briefly describe the three major steps involved in DNA fingerprinting using the Restriction Fragment Length Polymorphism (RFLP) technique.
Answer
1. Restriction digestion of genomic DNA, 2. Gel electrophoresis and Southern transfer, 3. Hybridisation with a labelled probe and detection of fragment patterns.
Step-by-step solution
- Step 1: Isolate genomic DNA from the individual and digest it with a specific restriction endonuclease to generate fragments of varying lengths.
- Step 2: Separate the fragments by agarose gel electrophoresis and transfer them onto a nylon membrane (Southern blot).
- Step 3: Hybridise the membrane with a labelled DNA probe that binds to a target sequence; visualise the bound probe to obtain a pattern of fragment sizes characteristic of the individual.
Common mistake: Omitting the hybridisation step or mixing up its order
Concept tested: RFLP fingerprinting steps
Published · How this solution was prepared and checked
← All Biotechnology Principles and Processes questions