Nucleic acid purification from plants, animals and microbes in under 30 seconds.
Nucleic acid purification from plants, animals and microbes in under 30 seconds.
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DOI：
10.1371/journal.pbio.2003916
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发表时间：
2017-11
期刊：
影响因子：
9.8
通讯作者：
Botella JR
中科院分区：
文献类型：
--
作者：
Zou Y;Mason MG;Wang Y;Wee E;Turni C;Blackall PJ;Trau M;Botella JR
核酸扩增是一种强大的分子生物学工具，尽管由于从生物样品中提取核酸所需的方法相对繁琐，其在现代实验室环境之外的使用受到限制。为了解决这个问题，我们研究了多种材料对于核酸捕获和纯化的适用性。我们在此报告，未经处理的纤维素纸可以在几秒钟内快速捕获核酸，并在单个洗涤步骤中保留它们，同时复杂生物样品中存在的污染物被快速去除。基于这些知识，我们成功创建了一种无需设备的核酸提取试纸方法，可以在不到 30 秒的时间内从困难的生物样本（例如成年树木的血液和叶子）中从植物、动物和微生物中获取可扩增的 DNA 和 RNA。这种方法的简单性和速度以及合适材料（例如普通纸巾）的低成本和可用性，意味着现在研究人员和更广泛的社区更容易获得和负担得起核酸提取。此外，当与等温扩增和裸眼 DNA 可视化技术的最新进展相结合时，试纸提取技术使得在资源有限的环境中（包括大学和高中教室、现场环境和发展中国家）可以进行分子诊断测定。事实证明，核酸扩增在世界各地的实验室中对于从诊断到基因分型的众多应用都是不可或缺的。任何旨在扩增 DNA 或 RNA 的应用的第一步都是从复杂的生物样品中提取核酸；这项任务传统上需要专门的设备、训练有素的技术人员和多个液体处理步骤。正是当前核酸分离方法的复杂性限制了许多 DNA 扩增技术在现代实验室环境之外的使用。因此，在本研究中，我们研究了简化核酸提取的新材料和方法。我们发现基于纤维素的滤纸可用于快速结合核酸，在短暂的洗涤步骤中保留它们以去除污染物，然后将它们直接洗脱到扩增反应中。然后，我们对纤维素过滤器进行了改造，制造了一种试纸，可用于在 30 秒内从各种植物、动物和微生物样品中纯化核酸，而不需要任何专门的设备。我们的方法的速度和简单性使其非常适合实验室内外基于核酸扩增的应用，包括资源有限的环境，例如偏远现场、发展中国家和教学机构。
Nucleic acid amplification is a powerful molecular biology tool, although its use outside the modern laboratory environment is limited due to the relatively cumbersome methods required to extract nucleic acids from biological samples. To address this issue, we investigated a variety of materials for their suitability for nucleic acid capture and purification. We report here that untreated cellulose-based paper can rapidly capture nucleic acids within seconds and retain them during a single washing step, while contaminants present in complex biological samples are quickly removed. Building on this knowledge, we have successfully created an equipment-free nucleic acid extraction dipstick methodology that can obtain amplification-ready DNA and RNA from plants, animals, and microbes from difficult biological samples such as blood and leaves from adult trees in less than 30 seconds. The simplicity and speed of this method as well as the low cost and availability of suitable materials (e.g., common paper towelling), means that nucleic acid extraction is now more accessible and affordable for researchers and the broader community. Furthermore, when combined with recent advancements in isothermal amplification and naked eye DNA visualization techniques, the dipstick extraction technology makes performing molecular diagnostic assays achievable in limited resource settings including university and high school classrooms, field-based environments, and developing countries. Nucleic acid amplification has proven to be indispensable in laboratories around the world for a myriad of applications from diagnostics to genotyping. The first step in any application aiming to amplify DNA or RNA is the extraction of nucleic acids from a complex biological sample; a task traditionally requiring specialised equipment, trained technicians, and multiple liquid handling steps. It is this complexity of current nucleic acid isolation methods that limit the use of many DNA amplification technologies outside of the modern laboratory environment. Therefore, in this study, we investigated new materials and approaches to simplify nucleic acid extraction. We found that cellulose-based filter paper can be used to rapidly bind nucleic acids, retain them during a short washing step to remove contaminants, and then elute them directly into the amplification reaction. We then adapted the cellulose filter to create a dipstick that can be used to purify nucleic acids from a wide range of plant, animal, and microbe samples in less than 30 seconds without the need for any specialised equipment. The speed and simplicity of our method makes it ideally suited for nucleic acid amplification-based applications both within and outside the laboratory, including limited resource settings such as remote field sites, developing countries, and teaching institutions.
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影响因子：
3.9
作者：
Linnes JC;Fan A;Rodriguez NM;Lemieux B;Kong H;Klapperich CM
通讯作者：
Klapperich CM
DOI：
10.1101/sqb.1968.033.01.033
发表时间：
1968-01-01
期刊：
COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY
影响因子：
--
作者：
ALBERTS, BM;AMODIO, FJ;FERRIS, FL
通讯作者：
FERRIS, FL
影响因子：
4.2
作者：
Kim, Jitae;Mauk, Michael;Bau, Haim H.
通讯作者：
Bau, Haim H.
影响因子：
2.2
作者：
Liesenfeld, O.;Lehman, L.;Kost, G.
通讯作者：
Kost, G.
影响因子：
5
作者：
Berensmeier S
通讯作者：
Berensmeier S