title: Fluorescence lifetime for on-the-fly multiplex detection of DNA restriction fragments in capillary electrophoresis.
abstract: This paper describes the first use of frequency-domain fluorescence lifetime for multiplex detection of DNA restriction fragments in capillary electrophoresis (CE). The fragments were labeled with monomeric intercalating dyes that can be excited by either the 488- or 514-nm line of an argon ion laser and have lifetimes in the range of 0.5-2.5 ns. We were able to achieve multiplex lifetime detection in the CE separation of a restriction fragment digest and a DNA size ladder in the same run, for fragments shorter than 700 bp. Different gel buffer systems, including a modified polyacrylamide gel and several tris-borate-EDTA/hydroxyethylcellulose (TBE/HEC) gels, were investigated for separation and detection of the dye-labeled DNA fragments. Best results for both electrophoretic resolution and lifetime detection were obtained using a gel containing 1% high molecular weight (90,000-105,000) HEC and 0.3% low molecular weight (24,000-27,000) HEC in TBE buffer.
authors.affiliation: Department of Chemistry, P. M. Gross Chemical Laboratory, Duke University, Durham, North Carolina 27708, USA.
authors.forename: S L
authors.initials: SL
authors.lastname: McIntosh
authors.forename: B K
authors.initials: BK
authors.lastname: Nunnally
authors.forename: A R
authors.initials: AR
authors.lastname: Nesbit
authors.forename: T G
authors.initials: TG
authors.lastname: Deligeorgiev
authors.forename: N I
authors.initials: NI
authors.lastname: Gadjev
authors.forename: L B
authors.initials: LB
authors.lastname: McGown
mesh_terms: D004247:DNA / Q000737:chemistry*; D019075:Electrophoresis, Capillary; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D007364:Intercalating Agents; D013056:Spectrophotometry, Ultraviolet
chemical_list: D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D007364:Intercalating Agents; D004247:DNA