By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX |
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Authors: | Fabian Tolle Julian Wilke Jesper Wengel Günter Mayer |
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Affiliation: | 1. LIMES Institute, University of Bonn, Bonn, Germany.; 2. Nucleic Acid Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense M, Denmark.; Imperial College London, United Kingdom, |
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Abstract: | The selection of nucleic acid aptamers is an increasingly important approach to generate specific ligands binding to virtually any molecule of choice. However, selection-inherent amplification procedures are prone to artificial by-product formation that prohibits the enrichment of target-recognizing aptamers. Little is known about the formation of such by-products when employing nucleic acid libraries as templates. We report on the formation of two different forms of by-products, named ladder- and non-ladder-type observed during repetitive amplification in the course of in vitro selection experiments. Based on sequence information and the amplification behaviour of defined enriched nucleic acid molecules we suppose a molecular mechanism through which these amplification by-products are built. Better understanding of these mechanisms might help to find solutions minimizing by-product formation and improving the success rate of aptamer selection. |
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