Biomaterial Database

Chromatographic separation of DNA restriction fragments. 1987

R Hecker, and D Riesner

Reversed-phase liquid chromatography (RPC-5) was the first chromatographic technique to be successfully applied to DNA restriction fragments. Size-exclusion high-performance liquid chromatography (HPLC) has the advantage of wide variability in buffer conditions but is restricted to an upper size limit at 800 base pairs. Anion-exchange HPLC is most versatile and may best be scaled up for the preparation of milligram amounts of specific DNA fragments. The preparation of the sample before and after chromatography is described. Because several applications of DNA fragments will be complemented by RNA fragments, their preparation and purification is included. Disposable small-scale columns are available for processing many samples. Future medical applications of HPLC of DNA fragments concern medical microbiology, human genetics and forensic medicine.

UI	MeSH Term	Description	Entries
D002845	Chromatography	Techniques used to separate mixtures of substances based on differences in the relative affinities of the substances for mobile and stationary phases. A mobile phase (fluid or gas) passes through a column containing a stationary phase of porous solid or liquid coated on a solid support. Usage is both analytical for small amounts and preparative for bulk amounts.	Chromatographies
D004262	DNA Restriction Enzymes	Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.	Restriction Endonucleases,DNA Restriction Enzyme,Restriction Endonuclease,Endonuclease, Restriction,Endonucleases, Restriction,Enzymes, DNA Restriction,Restriction Enzyme, DNA,Restriction Enzymes, DNA
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia