4.7 Review

ABCs of DNA aptamer and related assay development

Journal

BIOTECHNOLOGY ADVANCES
Volume 35, Issue 2, Pages 275-301

Publisher

PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.biotechadv.2017.01.003

Keywords

Aptamers; SELEX; Aptamer library; Colorimetric assay; Aptamer Linked Immunosorbent Assay (ALISA); FRET; ssDNA generation; Post-SELEX optimization; Translation of aptamers

Funding

  1. Department of Biotechnology, Govt of India
  2. Indian Council of Medical Research, Govt. of India

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This review is intended to guide the novice in aptamer research and development to understand virtually all of the aptamer development options and currently available assay modalities. Aptamer development topics range from discussions of basic and advanced versions of Systematic Evolution of Ligands by EXponential Enrichment (SELEX) and SELEX variations involving incorporation of exotic unnatural nucleotides to expand library diversity for even greater aptamer affinity and specificity to improved next generation methods of DNA sequencing, screening and tracking aptamer development throughout the SELEX process and characterization of lead aptamer candidates. Aptamer assay development topics include descriptions of various colorimetric and fluorescent assays in microplates or on membranes including homogeneous beacon and multiplexed Fluorescence Resonance Energy Transfer (FRET) assays. Finally, a discussion of the potential for marketing successful aptamer-based assays or test kits is included. (C) 2017 Elsevier Inc. All rights reserved.

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