Tournier Estelle, Amenc L., Pablo Anne-Laure, Legname E., Blanchart Eric, Plassard C., Robin A., 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, p. 182-191. ISSN 2215-0161.
Titre du document
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
Année de publication
2015
Auteurs
Tournier Estelle, Amenc L., Pablo Anne-Laure, Legname E., Blanchart Eric, Plassard C., Robin A., Bernard Laetitia
Source
MethodsX, 2015,
2, p. 182-191 ISSN 2215-0161
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 efficient on 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 from the previous tropical soils appeared to correlate better to soil respiration than DNA, which is positively influenced by soil clay content.
Plan de classement
Sciences fondamentales / Techniques d'analyse et de recherche [020]
;
Pédologie [068]
Description Géographique
MADAGASCAR ; ZONE TROPICALE
Localisation
Fonds IRD [F B010068850]
Identifiant IRD
fdi:010068850
