Modification of a commercial DNA extraction kit for safe and rapid recovery of DNA and RNA simultaneously from soil, without the use of harmful solvents
E. Tournier, L. Amenc, A.L. Pablo, E. Legname, E. Blanchart, C. Plassard, A. Robin, L. Bernard
TL;DR
This paper presents a modified DNA and RNA extraction method for soil that avoids harmful solvents and works well on challenging tropical soils.
Contribution
A solvent-free, simultaneous DNA and RNA extraction protocol for soil using commercial kits, optimized for tropical soils.
Findings
The protocol efficiently extracts nucleic acids from tropical soils with high clay and oxide content.
RNA recovery correlates better with soil respiration, while DNA recovery is influenced by clay content.
The method avoids harmful solvents and can be adapted for fungal parameter assessment.
Abstract
An optimized method, based on the coupling of two commercial kits, is described for the extraction of soil nucleic acids, with simultaneous extraction and purification of DNA and RNA following a cascade scheme and avoiding the use of harmful solvents. The protocol canmonitor the variations in the recovery yield of DNA and RNA from soils of various types.The quantitative version of the protocol was obtained by testing the starting soil quantity, the grinding parameters and the final elution volumes, in order to avoid saturation of both kits. •A first soil-crushing step in liquid nitrogen could be added for the assessment of fungal parameters.•The protocol was efficienton different tropical soils, including Andosol, while their high contents of clays, including poorly crystalline clays, and Fe and Al oxides usually make the nucleic acid extraction more difficult.•The RNA recovery yield…
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Taxonomy
TopicsGraphite, nuclear technology, radiation studies · Nuclear reactor physics and engineering · Iron and Steelmaking Processes