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使用一步微波辅助工艺在室温下实现DNA的长期稳定。

Long-term stabilization of DNA at room temperature using a one-step microwave assisted process.

作者信息

Narvaez Villarrubia Claudia W, Tumas Keyla C, Chauhan Rajat, MacDonald Thomas, Dattelbaum Andrew M, Omberg Kristin, Gupta Gautam

机构信息

Materials Physics and Applications Divison, Los Alamos National Laboratory, Los Alamos, NM USA.

Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM USA.

出版信息

Emergent Mater. 2022;5(2):307-314. doi: 10.1007/s42247-021-00208-3. Epub 2021 Mar 23.

Abstract

UNLABELLED

Long-term stabilization of DNA is needed for forensic, clinical, in-field operations and numerous other applications. Although freezing (<-20 °C) and dry storage are currently the preferential methods for long-term storage, a noticeable pre-analytical degradation of DNA over time, upfront capital investment and recurring costs have demonstrated a need for an alternative long-term room-temperature preservation method. Herein, we report a novel, fast (~5 min) silica sol-gel preparation method using a standard microwave-initiated polymerization reaction amenable to stabilization of DNA. The method involves use of one chemical, tetramethoxy silane (TMOS) and eliminates the use of alcohol as co-solvent and catalysts such as acids. In addition, the process involves minimal technical expertise, thus making it an ideal choice for resource-challenged countries and in-field applications. The sol-gel is capable to store and stabilize DNA in ambient conditions for 210 days. DNA recovered from the sol-gel showed no significant nucleolytic and/or oxidative degradation, outperforming conventional storage conditions at -20 °C, and reported state-of-the-art technology.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1007/s42247-021-00208-3.

摘要

未标注

法医、临床、现场操作及众多其他应用都需要DNA的长期稳定保存。尽管冷冻(<-20°C)和干燥储存目前是长期储存的首选方法,但随着时间的推移,DNA在分析前会出现明显降解,前期资本投资和经常性成本表明需要一种替代性的长期室温保存方法。在此,我们报告了一种新颖、快速(约5分钟)的硅溶胶-凝胶制备方法,该方法使用标准的微波引发聚合反应,适用于DNA的稳定化。该方法仅使用一种化学物质四甲氧基硅烷(TMOS),无需使用醇类作为共溶剂以及酸等催化剂。此外,该过程所需的技术专业知识极少,因此对于资源匮乏的国家和现场应用而言是理想之选。这种溶胶-凝胶能够在环境条件下储存和稳定DNA达210天。从溶胶-凝胶中回收的DNA未显示出明显的核酸降解和/或氧化降解,优于-20°C的传统储存条件以及已报道的最先进技术。

补充信息

在线版本包含可在10.1007/s42247-021-00208-3获取的补充材料。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cba/7986179/1f652673a409/42247_2021_208_Fig1_HTML.jpg

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