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A novel approach to imaging and visualization of minute amounts of DNA in small volume samples.
Ceresa, Luca; Kitchner, Emma; Seung, Michael; Bus, Magdalena M; Budowle, Bruce; Chavez, Jose; Gryczynski, Ignacy; Gryczynski, Zygmunt.
Afiliação
  • Ceresa L; Department of Physics and Astronomy, Texas Christian University, 2800 S. University, Dr. Fort Worth, Texas, 76129, USA. luca.ceresa@tcu.edu.
  • Kitchner E; Department of Physics and Astronomy, Texas Christian University, 2800 S. University, Dr. Fort Worth, Texas, 76129, USA. luca.ceresa@tcu.edu.
  • Seung M; Department of Physics and Astronomy, Texas Christian University, 2800 S. University, Dr. Fort Worth, Texas, 76129, USA. luca.ceresa@tcu.edu.
  • Bus MM; Center for Human Identification, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, CBH-250, Ft Worth, TX 76107, USA.
  • Budowle B; Department of Microbiology, Immunology and Genetics, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, Ft Worth, TX 76107, USA.
  • Chavez J; Center for Human Identification, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, CBH-250, Ft Worth, TX 76107, USA.
  • Gryczynski I; Department of Microbiology, Immunology and Genetics, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, Ft Worth, TX 76107, USA.
  • Gryczynski Z; Department of Physics and Astronomy, Texas Christian University, 2800 S. University, Dr. Fort Worth, Texas, 76129, USA. luca.ceresa@tcu.edu.
Analyst ; 146(21): 6520-6527, 2021 Oct 25.
Article em En | MEDLINE | ID: mdl-34559174
This report presents a novel approach for detecting and visualizing small to trace amounts of DNA in a sample. By utilizing both the change in emission spectrum and change in fluorescence lifetime, there is a significant increase in detection sensitivity allowing for the imaging/visualizing of a picograms amount of DNA in a microliters volume. As in the previous reports, one of the oldest DNA intercalators, Ethidium Bromide (EtBr), is employed as a model system. With this new approach, it is feasible to visualize just a few hundred picograms of DNA without the need for prior DNA amplification. The sensitivity can later be largely improved by using an intercalator that exhibits a higher affinity to DNA and a larger fluorescence change upon binding to DNA (e.g., ethidium homodimer, YOYO, or Diamond nucleic acid dyes).
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: DNA / Substâncias Intercalantes Idioma: En Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: DNA / Substâncias Intercalantes Idioma: En Ano de publicação: 2021 Tipo de documento: Article