Restriction Fragment Length Polymorphism (RFLP) analysis is based on two techniques that are widely used in modern molecular biology: the restriction endonuclease digestion of DNA and the transfer of DNA fragments to a filter, onto which can then be hybridized a labelled DNA fragment (1). Type II restriction endonucleases of bacteria recognize and cut specific nucleotide motifs in a DNA sequence (the enzymes commonly used for RFLP analysis recognize 4–6 base-pair sequences). They are, therefore, capable of reducing complex DNA, such as plant DNA, to a population of fragments with discrete sizes. In practice, fragments range in size from a few to more than several thousand base-pairs.
KeywordsRestriction Fragment Length Polymorphism High Specific Activity Restriction Fragment Length Polymorphism Analysis Restriction Fragment Length Polymorphism Marker Geiger Counter
Unable to display preview. Download preview PDF.
- 7.Bishop MD, Kappes SM, Keele JW, Stone RT, Sunden SL, Hawkins GA, Toldo SS, Fries R, Grosz MD, Yoo J et al. (1994) A genetic linkage map for cattle. Genetics 136: 619–639.Google Scholar
- 9.Burr B, Evola SV, Burr FA, Beckmann JS (1983) The application of restriction fragment length polymorphisms to plant breeding. In: Setlow JK, Hollaender A (eds) Genetic Engineering Principles and Methods, Vol 5. Plenum, New York, pp 45–59.Google Scholar
- 12.Medveczky P, Chang CW, Oste C, Mulder C (1987) Rapid vacuum driven transfer of DNA and RNA from gels to solid supports. BioTechniques 5: 24.Google Scholar
- 14.Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY; Cold Spring Harbor Laboratory.Google Scholar
- 17.Konat G, Laszkiewicz I, Bednarczuk T, Kanoh M, Wiggins RC (1991) Generation of radioactive and nonradioactive ssDNA hybridization probes by polymerase chain reaction. Technique 3 (2): 64–68.Google Scholar
- 18.Anderson MLM, Young BD (1987) Quantitative filter hybridisation. In: Harnes BD, Higgins SJ (ed) Nucleic acid hybridisation a practical approach. IRL press, Oxford UK, pp 73–111.Google Scholar