How does restriction fragment length polymorphism (RELP) technique work?
Posted June 4, 2024
Answer
In this technique:
- DNA is first cut into fragments using restriction endonuclease enzymes that recognize specific sequences.
- The DNA fragments are separated by size using agarose or polyacrylamide gel electrophoresis.
- The different-sized DNA fragments are denatured into single strands using sodium hydroxide (NaOH).
- The single-stranded DNA fragments are transferred onto a nitrocellulose membrane through electro-blotting or capillary blotting.
- The nitrocellulose membrane is fixed by autoclaving and then blocked with casein or bovine serum albumin to prevent non-specific binding.
- The labeled RFLP probe is hybridized with the complementary DNA on the membrane and the resulting bands are visualized using autoradiography.
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