A modular method for the extraction of DNA and RNA, and the separation of DNA pools from diverse environmental sample types

• Main Authors: Mark Alexander Lever, Andrea eTorti, Philip eEickenbusch, Alexander Bryce Michaud, Tina eŠantl-Temkiv, Bo Barker Jørgensen
• Format: Article
• Language: English
• Published: Frontiers Media S.A. 2015-05-01
• Series: Frontiers in Microbiology
• Subjects: DNA; RNA; extracellular; intracellular; modular; extraction
• Online Access: http://journal.frontiersin.org/Journal/10.3389/fmicb.2015.00476/full

A method for the extraction of nucleic acids from a wide range of environmental samples was developed. This method consists of several modules, which can be individually modified to maximize yields in extractions of DNA and RNA or separations of DNA pools. Modules were designed based on elaborate tests, in which permutations of all nucleic acid extraction steps were compared. The final modular protocol is suitable for extractions from igneous rock, air, water, and sediments. Sediments range from high-biomass, organic rich coastal samples to samples from the most oligotrophic region of the world’s oceans and the deepest borehole ever studied by scientific ocean drilling. Extraction yields of DNA and RNA are higher than with widely used commercial kits, indicating an advantage to optimizing extraction procedures to match specific sample characteristics. The ability to separate soluble extracellular DNA pools without cell lysis from intracellular and particle-complexed DNA pools may enable new insights into the cycling and preservation of DNA in environmental samples in the future. A general protocol is outlined, along with recommendations for optimizing this general protocol for specific sample types and research goals.