A decade of high-resolution liquid chromatography of nucleic acids on styrene-divinylbenzene copolymers.
The introduction of alkylated, nonporous poly-(styrene-divinylbenzene) microparticles in 1992 enabled the subsequent development of denaturing HPLC that has emerged as the most sensitive screening method for mutations to date. Denaturing HPLC has provided unprecedented insight into human origins and prehistoric migrations, accelerated the cloning of genes involved in mono- and polygenic traits, and facilitated the mutational analysis of more than a hundred candidate genes of human disease. A significant step toward increased sample-throughput and information content was accomplished by the recent introduction of monolithic poly(styrene-divinylbenzene) capillary columns. They have enabled the construction of capillary arrays amenable to multiplex analysis of fluorescent dye-labeled nucleic acids by laser-induced fluorescence detection. Hyphenation of denaturing HPLC with electrospray ionization mass spectrometry, on the other hand, has allowed the direct elucidation of the chemical nature of DNA variation and determination of phase of multiple alleles on a chromosome.
Oefner
Peter J
PJ
Genome Technology Center, Stanford University, 855 California Avenue, Palo Alto 94304, USA. oefner@genome.stanford.edu
Huber
Christian G
CG
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D009691:Nucleic Acid Denaturation; D009696:Nucleic Acids / Q000032:analysis*; D020058:Styrene / Q000737:chemistry*; D014753:Vinyl Compounds / Q000737:chemistry*
D009696:Nucleic Acids; D014753:Vinyl Compounds; D020058:Styrene; C004985:divinyl benzene