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Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay

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 in...

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Autores principales: Karbaschi, Mahsa, Ji, Yunhee, Mujawar, Mubarak A., Mendoza, Mario, Marquez, Juan S., Sonawane, Apurva, Shah, Pratikkumar, Ross, Chris, Bhansali, Shekhar, Cooke, Marcus S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10138556/
https://www.ncbi.nlm.nih.gov/pubmed/37108351
http://dx.doi.org/10.3390/ijms24087187
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author Karbaschi, Mahsa
Ji, Yunhee
Mujawar, Mubarak A.
Mendoza, Mario
Marquez, Juan S.
Sonawane, Apurva
Shah, Pratikkumar
Ross, Chris
Bhansali, Shekhar
Cooke, Marcus S.
author_facet Karbaschi, Mahsa
Ji, Yunhee
Mujawar, Mubarak A.
Mendoza, Mario
Marquez, Juan S.
Sonawane, Apurva
Shah, Pratikkumar
Ross, Chris
Bhansali, Shekhar
Cooke, Marcus S.
author_sort Karbaschi, Mahsa
collection PubMed
description 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.
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spelling pubmed-101385562023-04-28 Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay Karbaschi, Mahsa Ji, Yunhee Mujawar, Mubarak A. Mendoza, Mario Marquez, Juan S. Sonawane, Apurva Shah, Pratikkumar Ross, Chris Bhansali, Shekhar Cooke, Marcus S. Int J Mol Sci Brief Report 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. MDPI 2023-04-13 /pmc/articles/PMC10138556/ /pubmed/37108351 http://dx.doi.org/10.3390/ijms24087187 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Brief Report
Karbaschi, Mahsa
Ji, Yunhee
Mujawar, Mubarak A.
Mendoza, Mario
Marquez, Juan S.
Sonawane, Apurva
Shah, Pratikkumar
Ross, Chris
Bhansali, Shekhar
Cooke, Marcus S.
Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay
title Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay
title_full Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay
title_fullStr Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay
title_full_unstemmed Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay
title_short Development of a Novel, Automated High-Throughput Device for Performing the Comet Assay
title_sort development of a novel, automated high-throughput device for performing the comet assay
topic Brief Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10138556/
https://www.ncbi.nlm.nih.gov/pubmed/37108351
http://dx.doi.org/10.3390/ijms24087187
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