ArticleAnalytical biochemistry2022
Optimized quantitative PCR analysis of random DNA aptamer libraries.
Article in Analytical biochemistry, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 13 citations in OpenAlex.
- Article
- Cell-Free DNA-Based Theranostics for Inflammatory Disorders.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Review
- Immobilization-free SELEX for aptamer discovery targeting colorectal cancer-derived small extracellular vesicles.Journal of nanobiotechnology · 2025Article
- An Unbiased Cell-Culture Selection Yields DNA Aptamers as Novel Senescent Cell-Specific Reagents.Aging cell · 2025Article
- Methods for cell- and tissue-specific DNA aptamer selection.Molecular therapy. Methods & clinical development · 2025Article
- DNA aptamers that modulate biological activity of model neurons.Molecular therapy. Nucleic acids · 2024Article
- IMPATIENT-qPCR: monitoring SELEX success during in vitro aptamer evolution.Applied microbiology and biotechnology · 2024Article
- Peroxidase proximity selection to identify aptamers targeting a subcellular location.PNAS nexus · 2023Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
The quantitative polymerase chain reaction (qPCR) with detection of duplex DNA yield by intercalator fluorescence is a common and essential technique in nucleic acid analysis. We encountered unexpected results when applying standard qPCR methods to the quantitation of random DNA libraries flanked by regions of fixed sequence, a configuration essential for in vitro selection experiments. Here we describe the results of experiments revealing why conventional qPCR methods can fail to allow automated analysis in such cases, and simple solutions to this problem. In particular we show that renaturation of PCR products containing random regions is incomplete in late PCR cycles when extension fails due to reagent depletion. Intercalator fluorescence can then be lost at standard interrogation temperatures. We show that qPCR analysis of random DNA libraries can be achieved simply by adjusting the step at which intercalator fluorescence is monitored so that the yield of annealed constant regions is detected rather than the yield of full duplex DNA products.
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Registered trials
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