RFLP (Restriction Fragment Length Polymorphism)
Introduction to technique: Restriction Fragment Length Polymorphism, RFLP is a method of genetic analysis that allows individuals to be identified on the basis of unique patterns of restriction enzyme cutting in the particular regions of DNA. This technique takes an advantage of the polymorphisms occur in individual people's genetic codes. Even though all members of a particular specie have fundamentally the same genetic makeup, but these slight differences account for variations in phenotype between individuals.
Historical perspective: A British geneticist named Alec Jeffreys from Leicester University is accredited for the discovery and development of the RFLP method of DNA analysis and testing. He developed this method in 1985 as the
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A match may be confirmed if two samples have RFLP band sizes that are all within 5% of one another in size.
Figure 1: "RFLP analysis"
Applications of RFLP: Several genetic diseases can be detected by using RFLP analysis such as sickle cell anemia and cystic fibrosis. Specifically sickle cell anemia is caused by a single mutation of a nucleotide i.e. adenine is replaced by thymine. This mutation occurs at a point in the DNA sequence that is recognized by the restriction enzyme MstII in a person without the disease. RFLP of a person suffering from sickle cell disease have a single long band instead of two shorter bands because MstII cleavage will not likely to occur. Mutations in DNA between species are investigated by RFLP analysis. Figure 6:" Sickle cell anemia detected by RFLP analysis"
RFLPs can also be employed in different configurations to achieve various objectives:
• RFLPs can be used in criminal cases or paternity cases to determine the origin of a DNA sample (forensic