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

Tournier E., Amenc Laurie, Pablo Anne-laure, Legname Elvire, Blanchart Eric, Plassard Claude, Robin Agnès, Bernard Laetitia. 2015. 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. MethodsX, 2 : 182-191.

https://doi.org/10.1016/j.mex.2015.03.007

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Résumé : 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. o A first soil-crushing step in liquid nitrogen could be added for the assessment of fungal parameters. o 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. o The RNA recovery yield from the previous tropical soils appeared to correlate better to soil respiration than DNA, which is positively influenced by soil clay content.

Mots-clés Agrovoc : adn, arn, extraction par solvant, arn messager, purification

Mots-clés géographiques Agrovoc : France

Classification Agris : U30 - Méthodes de recherche
P34 - Biologie du sol
P33 - Chimie et physique du sol

Champ stratégique Cirad : Hors axes (2014-2018)

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