Biomaterial Database

Restriction fragment length polymorphism analysis. 2001

J Jarcho

Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA.

This unit describes Restriction Fragment Length Polymorphism (RFLP) analysis, which utilizes restriction endonuclease digestion to identify DNA sequence polymorphisms in genes or DNA regions of interest. When investigating families for inheritance of an RFLP, aliquots of genomic DNA from individual family members are digested to completion with the restriction enzyme known to generate the polymorphism of interest. After size fractionation on an agarose gel, DNA is transferred to a membrane by capillary action in a high-salt buffer. The gel is first treated with NaOH to denature DNA, and after neutralization, the gel is placed between buffer-soaked filter paper and a sheet of membrane. Labeled probe is hybridized overnight to the Southern blot. The blot is washed under conditions designed to remove all nonspecifically adherent probe and exposed to X-ray film. Identified fragment sizes differ among individuals and can be traced from generation to generation.

UI	MeSH Term	Description	Entries
D012150	Polymorphism, Restriction Fragment Length	Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment.	RFLP,Restriction Fragment Length Polymorphism,RFLPs,Restriction Fragment Length Polymorphisms
D004247	DNA	A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).	DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D004587	Electrophoresis, Agar Gel	Electrophoresis in which agar or agarose gel is used as the diffusion medium.	Electrophoresis, Agarose Gel,Agar Gel Electrophoresis,Agarose Gel Electrophoresis,Gel Electrophoresis, Agar,Gel Electrophoresis, Agarose
D005821	Genetic Techniques	Chromosomal, biochemical, intracellular, and other methods used in the study of genetics.	Genetic Technic,Genetic Technics,Genetic Technique,Technic, Genetic,Technics, Genetic,Technique, Genetic,Techniques, Genetic
D005826	Genetics, Medical	A subdiscipline of human genetics which entails the reliable prediction of certain human disorders as a function of the lineage and/or genetic makeup of an individual or of any two parents or potential parents.	Medical Genetics
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D015139	Blotting, Southern	A method (first developed by E.M. Southern) for detection of DNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.	Southern Blotting,Blot, Southern,Southern Blot