FluMag-SELEX as an advantageous method for DNA aptamer selection.
Aptamers are ssDNA or RNA oligonucleotides with very high affinity for their target. They bind to the target with high selectivity and specificity because of their specific three-dimensional shape. They are developed by the so-called Systematic Evolution of Ligands by Exponential Enrichment (SELEX) process. We have modified this method in two steps-use of fluorescent labels for DNA quantification and use of magnetic beads for target immobilization. Thus, radioactive labelling is avoided. Immobilization on magnetic beads enables easy handling, use of very small amounts of target for the aptamer selection, rapid and efficient separation of bound and unbound molecules, and stringent washing steps. We have called this modified SELEX technology FluMag-SELEX. With FluMag-SELEX we have provided a methodological background for our objective of being able to select DNA aptamers for targets with very different properties and size. These aptamers will be applied as new biosensor receptors. In this work selection of streptavidin-specific aptamers by FluMag-SELEX is described. The streptavidin-specific aptamers will be used to check the surface occupancy of streptavidin-coated magnetic beads with biotinylated molecules after immobilization procedures.
Stoltenburg
R
R
Department of Environmental Biotechnology, Centre for Environmental Research Leipzig-Halle GmbH (UFZ), Permoserstr. 15, 04318 Leipzig, Germany.
Reinemann
C
C
Strehlitz
B
B
D001665:Binding Sites; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D019921:Biotinylation; D020650:Combinatorial Chemistry Techniques; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D017931:DNA Primers / Q000032:analysis / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000032:analysis / Q000737:chemistry; D008024:Ligands; D008280:Magnetics; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000032:analysis* / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism
D017931:DNA Primers; D004277:DNA, Single-Stranded; D008024:Ligands; D009841:Oligonucleotides; D001710:Biotin; D004247:DNA; D019809:Streptavidin