Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay
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Published:2023-04-13
Issue:8
Volume:24
Page:7187
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ISSN:1422-0067
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Container-title:International Journal of Molecular Sciences
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language:en
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Short-container-title:IJMS
Author:
Karbaschi Mahsa1, Ji Yunhee2ORCID, Mujawar Mubarak A.3ORCID, Mendoza Mario3, Marquez Juan S.3ORCID, Sonawane Apurva3, Shah Pratikkumar3, Ross Chris4, Bhansali Shekhar3, Cooke Marcus S.2ORCID
Affiliation:
1. Independent Researcher, Sunnyvale, CA 94089, USA 2. Oxidative Stress Group, Department of Molecular Biosciences, University of South Florida, Tampa, FL 33620, USA 3. Department of Electrical and Computer Engineering, Florida International University, Miami, FL 33174, USA 4. Engineering Resources Group, Pembroke Pines, FL 33029, USA
Abstract
A comet assay is a trusted and widely used method for assessing DNA damage in individual eukaryotic cells. However, it is time-consuming and requires extensive monitoring and sample manipulation by the user. This limits the throughput of the assay, increases the risk of errors, and contributes to intra- and inter-laboratory variability. Here, we describe the development of a device which automates high throughput sample processing for a comet assay. This device is based upon our patented, high throughput, vertical comet assay electrophoresis tank, and incorporates our novel, patented combination of assay fluidics, temperature control, and a sliding electrophoresis tank to facilitate sample loading and removal. Additionally, we demonstrated that the automated device performs at least as well as our “manual” high throughput system, but with all the advantages of a fully “walkaway” device, such as a decreased need for human involvement and a decreased assay run time. Our automated device represents a valuable, high throughput approach for reliably assessing DNA damage with the minimal operator involvement, particularly if combined with the automated analysis of comets.
Funder
National Institution of Environmental Health Sciences of the National Institutes of Health
Subject
Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis
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