Abstract

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.

Details

Title
A high-throughput pipeline for scalable kit-free RNA extraction
Author
Han, Ping 1 ; Go, Maybelle K 1 ; Chow, Jeng Yeong 1 ; Xue, Bo 1 ; Lim, Yan Ping 1 ; Crone, Michael A 2 ; Storch Marko 3 ; Freemont Paul S 2 ; Yew Wen Shan 1   VIAFID ORCID Logo 

1  National University of Singapore, Synthetic Biology for Clinical and Technological Innovation, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431); Yong Loo Lin School of Medicine, National University of Singapore, Synthetic Biology Translational Research Programme, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431); Yong Loo Lin School of Medicine, National University of Singapore, Department of Biochemistry, Singapore, Singapore (GRID:grid.4280.e) (ISNI:0000 0001 2180 6431) 
2  Imperial College London, Section of Structural and Synthetic Biology, Department of Infectious Disease, Faculty of Medicine, London, UK (GRID:grid.7445.2) (ISNI:0000 0001 2113 8111); Imperial College White City Campus, London Biofoundry, Translation and Innovation Hub, London, UK (GRID:grid.7445.2) (ISNI:0000 0001 2113 8111); UK Dementia Research Institute Centre for Care Research and Technology, Based at Imperial College London and the, University of Surrey, London, UK (GRID:grid.5475.3) (ISNI:0000 0004 0407 4824) 
3  Imperial College London, Section of Structural and Synthetic Biology, Department of Infectious Disease, Faculty of Medicine, London, UK (GRID:grid.7445.2) (ISNI:0000 0001 2113 8111); Imperial College White City Campus, London Biofoundry, Translation and Innovation Hub, London, UK (GRID:grid.7445.2) (ISNI:0000 0001 2113 8111) 
Publication year
2021
Publication date
2021
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41598-021-02742-w
ProQuest document ID
2604985932
Copyright
© The Author(s) 2021. corrected publication 2021. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.