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A high-throughput pipeline for scalable kit-free RNA extraction

An overreliance on commercial, kit-based RNA extraction in the molecular diagnoses of infectious disease presents a challenge in the event of supply chain disruptions and can potentially hinder testing capacity in times of need. In this study, we adapted a well-established, robust TRIzol-based RNA e...

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Autores principales: Han, Ping, Go, Maybelle K., Chow, Jeng Yeong, Xue, Bo, Lim, Yan Ping, Crone, Michael A., Storch, Marko, Freemont, Paul S., Yew, Wen Shan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8636496/
https://www.ncbi.nlm.nih.gov/pubmed/34853385
http://dx.doi.org/10.1038/s41598-021-02742-w
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author Han, Ping
Go, Maybelle K.
Chow, Jeng Yeong
Xue, Bo
Lim, Yan Ping
Crone, Michael A.
Storch, Marko
Freemont, Paul S.
Yew, Wen Shan
author_facet Han, Ping
Go, Maybelle K.
Chow, Jeng Yeong
Xue, Bo
Lim, Yan Ping
Crone, Michael A.
Storch, Marko
Freemont, Paul S.
Yew, Wen Shan
author_sort Han, Ping
collection PubMed
description An overreliance on commercial, kit-based RNA extraction in the molecular diagnoses of infectious disease presents a challenge in the event of supply chain disruptions and can potentially hinder testing capacity in times of need. In this study, we adapted a well-established, robust TRIzol-based RNA extraction protocol into a high-throughput format through miniaturization and automation. The workflow was validated by RT-qPCR assay for SARS-CoV-2 detection to illustrate its scalability without interference to downstream diagnostic sensitivity and accuracy. This semi-automated, kit-free approach offers a versatile alternative to prevailing integrated solid-phase RNA extraction proprietary systems, with the added advantage of improved cost-effectiveness for high volume acquisition of quality RNA whether for use in clinical diagnoses or for diverse molecular applications.
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spelling pubmed-86364962021-12-03 A high-throughput pipeline for scalable kit-free RNA extraction Han, Ping Go, Maybelle K. Chow, Jeng Yeong Xue, Bo Lim, Yan Ping Crone, Michael A. Storch, Marko Freemont, Paul S. Yew, Wen Shan Sci Rep Article An overreliance on commercial, kit-based RNA extraction in the molecular diagnoses of infectious disease presents a challenge in the event of supply chain disruptions and can potentially hinder testing capacity in times of need. In this study, we adapted a well-established, robust TRIzol-based RNA extraction protocol into a high-throughput format through miniaturization and automation. The workflow was validated by RT-qPCR assay for SARS-CoV-2 detection to illustrate its scalability without interference to downstream diagnostic sensitivity and accuracy. This semi-automated, kit-free approach offers a versatile alternative to prevailing integrated solid-phase RNA extraction proprietary systems, with the added advantage of improved cost-effectiveness for high volume acquisition of quality RNA whether for use in clinical diagnoses or for diverse molecular applications. Nature Publishing Group UK 2021-12-01 /pmc/articles/PMC8636496/ /pubmed/34853385 http://dx.doi.org/10.1038/s41598-021-02742-w Text en © The Author(s) 2021, corrected publication 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Han, Ping
Go, Maybelle K.
Chow, Jeng Yeong
Xue, Bo
Lim, Yan Ping
Crone, Michael A.
Storch, Marko
Freemont, Paul S.
Yew, Wen Shan
A high-throughput pipeline for scalable kit-free RNA extraction
title A high-throughput pipeline for scalable kit-free RNA extraction
title_full A high-throughput pipeline for scalable kit-free RNA extraction
title_fullStr A high-throughput pipeline for scalable kit-free RNA extraction
title_full_unstemmed A high-throughput pipeline for scalable kit-free RNA extraction
title_short A high-throughput pipeline for scalable kit-free RNA extraction
title_sort high-throughput pipeline for scalable kit-free rna extraction
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8636496/
https://www.ncbi.nlm.nih.gov/pubmed/34853385
http://dx.doi.org/10.1038/s41598-021-02742-w
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