title: Just in time-selection: A rapid semiautomated SELEX of DNA aptamers using magnetic separation and BEAMing.
abstract: A semiautomated two-step method for in vitro selection of DNA aptamers using magnetic separation and solid-phase emulsion polymerase chain reaction has been developed. The application of a magnetic separator allows the simultaneous processing of up to 12 SELEXs (systematic evolution of ligands by exponential enrichment) with different targets or buffer conditions. Using a magnetic separator and covalent target immobilization on magnetic beads, the selection process was simplified and the substeps of aptamer/target incubation, washing, and elution of the aptamers were merged into one automated procedure called "FISHing". Without further processing the resulting FISHing eluates are suitable for BEAMing (beads, emulsion, amplification, and magnetics), which includes the amplification by emPCR (emulsion polymerase chain reaction) and strand separation by the implementation of covalently immobilized reverse primers on magnetic beads. The novel selection process has been proved and validated by selecting and characterization of aptamers to the wine fining agent lysozyme.
authors.affiliation: Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg , Grindelallee 117, 20146 Hamburg, Germany.
authors.forename: Tim
authors.initials: T
authors.lastname: Hünniger
authors.forename: Hauke
authors.initials: H
authors.lastname: Wessels
authors.forename: Christin
authors.initials: C
authors.lastname: Fischer
authors.forename: Angelika
authors.initials: A
authors.lastname: Paschke-Kratzin
authors.forename: Markus
authors.initials: M
authors.lastname: Fischer
mesh_terms: D052157:Aptamers, Nucleotide / Q000032:analysis*; D001331:Automation; D008280:Magnetics; D016133:Polymerase Chain Reaction / Q000379:methods; D052156:SELEX Aptamer Technique / Q000379:methods*; D020349:Surface Plasmon Resonance
chemical_list: D052157:Aptamers, Nucleotide