Rapid (30-second), equipment-free purification of nucleic acids using easy-to-make dipsticks

被引:64
作者
Mason, Michael G. [1 ]
Botella, Jose R. [1 ]
机构
[1] Univ Queensland, Sch Agr & Food Sci, Plant Genet Engn Lab, St Lucia, Qld, Australia
关键词
MEDIATED ISOTHERMAL AMPLIFICATION; DNA EXTRACTION; PCR;
D O I
10.1038/s41596-020-0392-7
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The complexity of current nucleic acid isolation methods limits their use outside of the modern laboratory environment. Here, we describe a fast and affordable method to purify nucleic acids from animal, plant, viral and microbial samples using a cellulose-based dipstick. Nucleic acids can be purified by dipping in-house-made dipsticks into just three solutions: the extract (to bind the nucleic acids), a wash buffer (to remove impurities) and the amplification reaction (to elute the nucleic acids). The speed and simplicity of this method make it ideally suited for molecular applications, both within and outside the laboratory, including limited-resource settings such as remote field sites and teaching institutions. Detailed instructions for how to easily manufacture large numbers of dipsticks in house are provided. Using the instructions, readers can create more than 200 dipsticks in <30 min and perform dipstick-based nucleic acid purifications in 30 s. The authors describe how to easily prepare a large number of dipsticks from cellulose-based filter paper and use them to rapidly purify nucleic acids from a variety of sources.
引用
收藏
页码:3663 / 3677
页数:17
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