Capillary electrophoresis of DNA for molecular diagnostics.
A number of applications of capillary zone electrophoresis (CZE) in sieving liquid polymers (notably linear polyacrylamides and cellulose) for the analysis of polymerase chain reaction (PCR) products of clinically relevant, diagnostic DNA, are reviewed. The fields covered are: human genetics, quantitative gene dosage, microbiology and virology, forensic medicine and therapeutic DNA (notably, antisense nucleotides). Some unique, novel developments are highlighted, such as: (i) nonisocratic CZE, i.e., temperature-programmed CZE for detection of DNA point mutations; (ii) the synthesis of novel N-substituted acrylamides, offering extreme resistance to alkaline hydrolysis coupled to high hydrophilicity. In the field of denaturing gradient gel electrophoresis (DGGE), as routinely performed in gel slabs, a novel methodology is described in CZE: double-gradient DGGE. In this technique, two gradients are simultaneously applied along the migration direction: a chemical (or thermal) denaturing gradient, for partially unwinding homo- and hetero-duplexes of DNA, and a porosity gradient, for recompacting diffuse bands melting over a broader range of denaturing conditions. It is thus demonstrated that chemical gradients, in addition to temperature gradients, can be easily implemented even in a capillary format.
Righetti
P G
PG
University of Verona, Department of Agricultural and Industrial Biotechnologies, Italy. righetti@imiucca.csi.unimi.it
Gelfi
C
C
D001424:Bacterial Infections / Q000175:diagnosis; D004247:DNA / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000639:trends; D005544:Forecasting; D006801:Humans; D014777:Virus Diseases / Q000175:diagnosis
D004247:DNA