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在硅胶溶胶 - 凝胶基质中于室温和高温下长期储存微小核糖核酸(miRNA)

Long term storage of miRNA at room and elevated temperatures in a silica sol-gel matrix.

作者信息

Chauhan Rajat, Kalbfleisch Theodore S, Potnis Chinmay S, Bansal Meenakshi, Linder Mark W, Keynton Robert S, Gupta Gautam

机构信息

Department of Chemical Engineering, University of Louisville Louisville Kentucky 40292 USA

Department of Chemistry, University of Louisville Louisville Kentucky 40292 USA.

出版信息

RSC Adv. 2021 Sep 23;11(50):31505-31510. doi: 10.1039/d1ra04719a. eCollection 2021 Sep 21.

Abstract

Storage of biospecimens in their near native environment at room temperature can have a transformative global impact, however, this remains an arduous challenge to date due to the rapid degradation of biospecimens over time. Currently, most isolated biospecimens are refrigerated for short-term storage and frozen (-20 °C, -80 °C, liquid nitrogen) for long-term storage. Recent advances in room temperature storage of purified biomolecules utilize anhydrobiosis. However, a near aqueous storage solution that can preserve the biospecimen nearly "as is" has not yet been achieved by any current technology. Here, we demonstrate an aqueous silica sol-gel matrix for optimized storage of biospecimens. Our technique is facile, reproducible, and has previously demonstrated stabilization of DNA and proteins, within a few minutes using a standard benchtop microwave. Herein, we demonstrate complete integrity of miRNA 21, a highly sensitive molecule at 4, 25, and 40 °C over a period of ∼3 months. In contrast, the control samples completely degrade in less than 1 week. We attribute excellent stability to entrapment of miRNA within silica-gel matrices.

摘要

在室温下将生物样本保存在其接近天然的环境中可能会对全球产生变革性影响,然而,由于生物样本会随时间快速降解,这一目标至今仍是一项艰巨的挑战。目前,大多数分离出的生物样本短期冷藏保存,长期则冷冻保存(-20℃、-80℃、液氮)。纯化生物分子的室温储存方面的最新进展利用了隐生现象。然而,目前尚无任何技术能实现一种近乎能原样保存生物样本的近水性储存溶液。在此,我们展示了一种用于优化生物样本储存的水性二氧化硅溶胶-凝胶基质。我们的技术简便、可重复,并且此前已证明使用标准台式微波炉在几分钟内就能实现DNA和蛋白质的稳定。在此,我们证明了miRNA 21在4℃、25℃和40℃下约3个月内的完全完整性。相比之下,对照样本在不到1周的时间内就完全降解了。我们将其优异的稳定性归因于miRNA被包裹在硅胶基质中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e0e7/9041656/fb58f5ea6232/d1ra04719a-f1.jpg

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