A novel approach to imaging and visualization of minute amounts of DNA in small volume samples

Luca Ceresa; Emma Kitchner; Michael Seung; Magdalena M. Bus; Bruce Budowle; Jose Chavez; Ignacy Gryczynski; Zygmunt Gryczynski

doi:10.1039/D1AN01391B

A novel approach to imaging and visualization of minute amounts of DNA in small volume samples

Luca Ceresa,

*^a Emma Kitchner,

^a Michael Seung,^a Magdalena M. Bus,^bc Bruce Budowle,^bc Jose Chavez,

^a Ignacy Gryczynski^a and Zygmunt Gryczynski^a

Author affiliations

* Corresponding authors

^a Department of Physics and Astronomy, Texas Christian University, 2800 S. University, Dr. Fort Worth, Texas, USA
E-mail: luca.ceresa@tcu.edu

^b Center for Human Identification, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, Ft Worth, TX 76107, USA

^c Department of Microbiology, Immunology and Genetics, Graduate School of Biomedical Sciences, University of North Texas Health Science Center, Ft Worth, TX 76107, USA

Abstract

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).

Article information

https://doi.org/10.1039/D1AN01391B

Article type

Paper

Submitted

02 Aug 2021

Accepted

08 Sep 2021

First published

20 Sep 2021

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Analyst, 2021,146, 6520-6527

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A novel approach to imaging and visualization of minute amounts of DNA in small volume samples

L. Ceresa, E. Kitchner, M. Seung, M. M. Bus, B. Budowle, J. Chavez, I. Gryczynski and Z. Gryczynski, Analyst, 2021, 146, 6520 DOI: 10.1039/D1AN01391B

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A novel approach to imaging and visualization of minute amounts of DNA in small volume samples

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