Journal Article

Nucleic acid purification from plants, animals and microbes in under 30 seconds

Trau, Matt, Botella, Jose Ramon, Wang, Yuling, Turni, Conny, Mason, Michael Glenn, Zou, Yiping, Wee, Eugene, Blackall, Patrick J.

2017

Overview

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.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Amplification / Animals / Arabidopsis - genetics / Authorship / Bacteria - genetics / Bioengineering / Biological properties / Biological samples / Biology and Life Sciences / Cellulose / Cellulose - chemistry / Contaminants / Deoxyribonucleic acid / Developing countries / Diagnostic systems / DNA / DNA - analysis / DNA - isolation & purification / Enzymes / Flowers & plants / Food / Funding / Genetic engineering / Genetic Techniques / Humans / Inventors / Laboratories / LDCs / Leaves / Methods / Methods and Resources / Microorganisms / Molecular biology / Nanoparticles / Nanotechnology / Nucleic Acid Amplification Techniques / Nucleic acids / Nucleic Acids - analysis / Nucleic Acids - isolation & purification / Oligonucleotides - genetics / Physical Sciences / Plant Leaves - genetics / Plants (botany) / Plants - genetics / Purification / Research and Analysis Methods / Ribonucleic acid / RNA / RNA - analysis / RNA - isolation & purification / Separation / Studies / Supervision / Visualization