Digital Polymerase Chain Reaction Paired with High-Speed Atomic Force Microscopy for Quantitation and Length Analysis of DNA Length Polymorphisms
| العنوان: | Digital Polymerase Chain Reaction Paired with High-Speed Atomic Force Microscopy for Quantitation and Length Analysis of DNA Length Polymorphisms |
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| المؤلفون: | A. L. Mikheikin, Jason Reed, Amir A. Toor, Sean Koebley, Alden Chesney, Loren Picco, Daniel Guest, Taha Al Juhaishi, Kevin Leslie, Xiaojie Zhang, Catherine H. Roberts, Wendy McConnell-Wells, Joshua H Lehman |
| المصدر: | ACS Nano. 14:15385-15393 |
| بيانات النشر: | American Chemical Society (ACS), 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Poor prognosis, Atomic force microscopy, General Engineering, General Physics and Astronomy, DNA, Sequence Analysis, DNA, Variant allele, Computational biology, Biology, Amplicon, Microscopy, Atomic Force, Polymerase Chain Reaction, Leukemia, Myeloid, Acute, chemistry.chemical_compound, Gene Frequency, fms-Like Tyrosine Kinase 3, chemistry, Humans, General Materials Science, Digital polymerase chain reaction, Flt3 gene |
| الوصف: | DNA length polymorphisms are found in many serious diseases, and assessment of their length and abundance is often critical for accurate diagnosis. However, measuring their length and frequency in a mostly wild-type background, as occurs in many situations, remains challenging due to their variable and repetitive nature. To overcome these hurdles, we combined two powerful techniques, digital polymerase chain reaction (dPCR) and high-speed atomic force microscopy (HSAFM), to create a simple, rapid, and flexible method for quantifying both the size and proportion of DNA length polymorphisms. In our approach, individual amplicons from each dPCR partition are imaged and sized directly. We focused on internal tandem duplications (ITDs) located within the FLT3 gene, which are associated with acute myeloid leukemia and often indicative of a poor prognosis. In an analysis of over 1.5 million HSAFM-imaged amplicons from cell line and clinical samples containing FLT3-ITDs, dPCR-HSAFM returned the expected variant length and variant allele frequency, down to 5% variant samples. As a high-throughput method with single-molecule resolution, dPCR-HSAFM thus represents an advance in HSAFM analysis and a powerful tool for the diagnosis of length polymorphisms. |
| تدمد: | 1936-086X 1936-0851 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::264499a268db25b47768fe45ef945d2c https://doi.org/10.1021/acsnano.0c05897 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....264499a268db25b47768fe45ef945d2c |
| قاعدة البيانات: | OpenAIRE |
Find this issue from the American Chemical Society | تدمد: | 1936086X 19360851 |
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