Version 1
: Received: 26 April 2024 / Approved: 26 April 2024 / Online: 26 April 2024 (19:59:38 CEST)
How to cite:
Daum, L.T.; Rodriguez, J.D.; Chambers, J.P. Development of Aptamers for RNase Inactivation in Xtract-Free™ Sample Collection and Transport Medium. Preprints2024, 2024041788. https://doi.org/10.20944/preprints202404.1788.v1
Daum, L.T.; Rodriguez, J.D.; Chambers, J.P. Development of Aptamers for RNase Inactivation in Xtract-Free™ Sample Collection and Transport Medium. Preprints 2024, 2024041788. https://doi.org/10.20944/preprints202404.1788.v1
Daum, L.T.; Rodriguez, J.D.; Chambers, J.P. Development of Aptamers for RNase Inactivation in Xtract-Free™ Sample Collection and Transport Medium. Preprints2024, 2024041788. https://doi.org/10.20944/preprints202404.1788.v1
APA Style
Daum, L.T., Rodriguez, J.D., & Chambers, J.P. (2024). Development of Aptamers for RNase Inactivation in Xtract-Free™ Sample Collection and Transport Medium. Preprints. https://doi.org/10.20944/preprints202404.1788.v1
Chicago/Turabian Style
Daum, L.T., John David Rodriguez and James P Chambers. 2024 "Development of Aptamers for RNase Inactivation in Xtract-Free™ Sample Collection and Transport Medium" Preprints. https://doi.org/10.20944/preprints202404.1788.v1
Abstract
There is significant need for development of a new environmentally friendly, extraction-free sample collection medium that can effectively preserve and protect genetic material for point-of-care, and/or self-collection, home-collection, and mail-back testing. Systematic Evolution of Ligands by Exponential Enrichment (SELEX) was used to create anti-ribonuclease (RNase) Deoxyribonucleic acid (DNA) aptamers against purified RNase A conjugated to paramagnetic carboxylated beads. Following 8 rounds of SELEX carried out under various stringency conditions, e.g., selection using Xtract-Free™ (XF) specimen collection medium, and elevated ambient temperature of 28 ºC, a panel of 5 aptamers was chosen following bioinformatic analysis using next-generation sequencing. The efficacy of aptamer inactivation of RNase was assessed by monitoring Ribonucleic acid (RNA) integrity by fluorometric and real-time RT-PCR analysis. Inclusion of aptamers in reaction incubations resulted in an 8,800 to 11,200-fold reduction of RNase activity, i.e., digestion of viral RNA compared to control. Thus, anti-RNase aptamers integrated into XF collection medium as well as other commercial reagents and kits have great potential for ensuring quality intact RNA for subsequent genomic analysis.
Keywords
Xtract-Fee™ medium; sample collection; qPCR; diagnostics; extraction-free detection; NAT; point of care; home-collect; self-collect; DNA aptamers; RNase; nucleases; 3rd generation specimen collection; AptaGuard
Subject
Biology and Life Sciences, Biology and Biotechnology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
