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Monitoring the progression of the in vitro selection of nucleic acid aptamers by denaturing high-performance liquid chromatography

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Abstract

Monitoring of in-vitro selection experiments is crucial in evaluation of the success and outcome of such approaches. Furthermore, monitoring running in parallel with the selection procedure enables early intervention and adjustment of stringency to achieve the desired activities of the selected nucleic acid species. Here we describe the use of a non-radioactive method that enables monitoring of a SELEX procedure on the basis of sequence diversity. We employ denaturing HPLC and describe for the first time an experimental set-up that is useful both for analysis of the progression of in-vitro selection experiments and for separation of distinct aptamer sequences.

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Acknowledgement

This work was supported by the DFG (Ma 3442/1-1) and the Fonds der Chemischen Industrie. The authors would like to thank Judith Junen for expert technical assistance.

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Authors and Affiliations

  1. Institute of Experimental Haematology and Transfusion Medicine, University of Bonn, Sigmund-Freud-Str. 25, 53127, Bonn, Germany

    Jens Müller, Osman El-Maarri, Johannes Oldenburg & Bernd Pötzsch

  2. Life and Medical Sciences Bonn, c/o Kekulé-Institute for Org. Chemistry and Biochemistry, University of Bonn, Gerhard-Domagk-Str. 1, 53121, Bonn, Germany

    Günter Mayer

Authors
  1. Jens Müller
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  2. Osman El-Maarri
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  3. Johannes Oldenburg
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  4. Bernd Pötzsch
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  5. Günter Mayer
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Correspondence to Günter Mayer.

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Müller, J., El-Maarri, O., Oldenburg, J. et al. Monitoring the progression of the in vitro selection of nucleic acid aptamers by denaturing high-performance liquid chromatography. Anal Bioanal Chem 390, 1033–1037 (2008). https://doi.org/10.1007/s00216-007-1699-8

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  • DOI: https://doi.org/10.1007/s00216-007-1699-8

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