title: Recovery of functional DNA inserts by electroendosmotic elution during gel electrophoresis.
abstract: In contrast to all previous preparative electrophoresis apparatus which used a pump, electroendosmotic elution uses bound electrical charges at the end of the separating gel to generate a buffer flow. The electroendosmotic flow increased with increasing currents and decreasing buffer concentrations: its exact characteristics for the built apparatus were determined. The electroendosmotic device was able to separate two DNA fragments differing in size by only 5% with a recovery over 95%. As demonstrated in practical examples of recovery and uses of DNA inserts, up to 10 micrograms of DNA per band can be loaded at a time. The recovered DNA can be used directly for nick-translation, ligation... without further treatment. The performances of the method are expected to improve still further if the charge density and pores of the electroendosmotic medium can be "made-to-order" to provide a better flow profile of the eluting buffer.
authors.affiliation: INSERM U15, Paris, France.
authors.forename: H V
authors.initials: HV
authors.lastname: Tan
authors.forename: A
authors.initials: A
authors.lastname: Kitzis
authors.forename: T
authors.initials: T
authors.lastname: Berthollet
authors.forename: G
authors.initials: G
authors.lastname: Hamard
authors.forename: C
authors.initials: C
authors.lastname: Beldjord
authors.forename: R
authors.initials: R
authors.lastname: Benarous
mesh_terms: D002021:Buffers; D004247:DNA / Q000302:isolation & purification*; D004262:DNA Restriction Enzymes; D004586:Electrophoresis / Q000295:instrumentation*; D004587:Electrophoresis, Agar Gel / Q000295:instrumentation*; D009995:Osmosis
chemical_list: D002021:Buffers; D004247:DNA; D004262:DNA Restriction Enzymes