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Application of the Open qPCR Instrument for the in Vitro Selection of DNA Aptamers against Epidermal Growth Factor Receptor and Drosophila C Virus.
Damase, Tulsi Ram; Miura, Tanya A; Parent, Christine E; Allen, Peter B.
Afiliación
  • Damase TR; University of Idaho , Department of Chemistry, 875 Perimeter Drive, Moscow, Idaho 83844-2343, United States.
  • Miura TA; University of Idaho , Department of Biological Sciences, 875 Perimeter Drive, MS 3051, Moscow, Idaho 83844-3051, United States.
  • Parent CE; University of Idaho , Department of Biological Sciences, 875 Perimeter Drive, MS 3051, Moscow, Idaho 83844-3051, United States.
  • Allen PB; University of Idaho , Department of Chemistry, 875 Perimeter Drive, Moscow, Idaho 83844-2343, United States.
ACS Comb Sci ; 20(2): 45-54, 2018 02 12.
Article en En | MEDLINE | ID: mdl-29293309
ABSTRACT
The low-cost Open qPCR instrument can be used for different tasks in the aptamer selection process quantification of DNA, cycle course optimization, screening, and final binding characterization. We have selected aptamers against whole Drosophila C virus (DCV) particles and recombinant epidermal growth factor receptor (EGFR). We performed systematic evolution of ligands by exponential enrichment (SELEX) using the Open qPCR to optimize each amplification step. The Open qPCR instrument identified the best aptamer candidate. The Open qPCR has the capacity to perform melt curves, and we used this function to perform thermofluorimetric analysis (TFA) to quantify target-aptamer binding. We confirmed target-aptamer binding using flow cytometry. A sandwich type luminescence bioassay based on our anti-DCV aptamer was sensitive to DCV and did not respond to a related virus, demonstrating that our selected anti-DCV aptamer can be used to specifically detect DCV.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Reacción en Cadena de la Polimerasa / Aptámeros de Nucleótidos / Dicistroviridae / Receptores ErbB Tipo de estudio: Prognostic_studies Idioma: En Revista: ACS Comb Sci Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Reacción en Cadena de la Polimerasa / Aptámeros de Nucleótidos / Dicistroviridae / Receptores ErbB Tipo de estudio: Prognostic_studies Idioma: En Revista: ACS Comb Sci Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos