title: DNA fragment separations by on-line combination of capillary isotachophoresis-capillary zone electrophoresis with UV detection.
abstract: A high-speed DNA fragment separation system based on an on-line combination of capillary ITP with CZE (CITP-CZE) and using UV detection at 260 nm was developed. A novel CITP-CZE buffer system of pH 6.1 was designed for the separation of ten DNA fragments with sizes ranging from 100 to 1000 bp. An effect of underivatized α-, β- and γ-cyclodextrins on the resolution of DNA fragments in the CZE step of the CITP-CZE combination was systematically investigated. Methylhydroxyethylcellulose present in the BGE was used to eliminate the EOF. DNA ladder fragments were separated within 10 min with LODs in the range of 1-5 ng/μL (S/N = 3). The RSDs of the migration time and peak area of individual DNA fragments were in the range of 1-3 and 3-9%, respectively. The developed CITP-CZE system was further applied to the analysis of digest plasmid DNA samples.
authors: Department of Molecular Biology, Faculty of Natural Sciences, Comenius University in Bratislava, Bratislava, Slovakia.
authors: Milan
authors: M
authors: Fraňo
authors: Department of Analytical Chemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Bratislava, Slovakia.
authors: Katarína
authors: K
authors: Džuganová
authors: Department of Organic Chemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Bratislava, Slovakia.
authors: Pavol
authors: P
authors: Koiš
authors: Department of Analytical Chemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Bratislava, Slovakia.
authors: Marián
authors: M
authors: Masár
mesh_terms: D004247:DNA / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D057625:Isotachophoresis / Q000379:methods*; D057230:Limit of Detection; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
chemical_list: D004247:DNA
keywords: Capillary isotachophoresis-capillary zone electrophoresis; DNA separation; Native cyclodextrins; UV detection