Restriction fragment

Restriction Fragment Length Polymorphism (RFLP) is a technique in molecular biology that exploits variations in homologous DNA sequences. It refers to a difference between samples of homologous DNA molecules from differing locations of restriction enzyme sites, and to a related laboratory technique by which these segments can be illustrated.

History

RFLP was first described in 1980 by David Botstein, Ronald J. Davis, Mark Skolnick, and Ray White. It was the first technique used in genetic mapping and genomic mapping, and was the basis for the first human genome maps.

Method

In RFLP, DNA is extracted and then digested by restriction enzymes. The resulting restriction fragments are separated according to their lengths by gel electrophoresis. The fragments are then transferred to a membrane through a blotting technique, and hybridized with a DNA probe that is complementary to the desired sequence. The probe is labeled with a radioactive or chemical tag.

Applications

RFLP has been used in forensic science to identify individuals or samples. It has also been used in gene mapping, hereditary disease diagnosis, and species differentiation.

Limitations

RFLP analysis is laborious and requires a large amount of DNA, which can limit its usefulness. It is also less sensitive than other techniques such as PCR.

See also

• DNA sequencing
• Genetic fingerprinting
• Molecular marker
• Polymerase chain reaction

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