The substitution of a T for an A in the protein coding sequence of the globin ge

Question

The substitution of a T for an A in the protein coding sequence of the globin gene introduced a SNP. This SNP changed an amino acid resulting in sickle cell anemia. Unaffected individuals have the DNA sequence GAGGAG, a BseRI restriction enzyme site, in their gene generating two DNA fragments of 2 and 4 kb. The A to T substitution occurs within the BseR1 site thus inactivating the site (GTGGAG). Draw the RFLP gel including results for a carrier, as well as homozygous normal and affected individuals.

Explanation / Answer

Restriction Fragment Length Polymorphism (RFLP) is a difference in homologous DNA sequences that can be detected by the presence of fragments of different lengths after digestion of the DNA samples in question with specific restriction endonucleases.RFLP, as a molecular marker, is specific to a single clone/restriction enzyme combination. An RFLP probe is a labeled DNA sequence that hybridizes with one or more fragments of the digested DNA sample after they were separated by gel electrophoresis, thus revealing a unique blotting pattern characteristic to a specific genotype at a specific locus.

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