Method for rapid DNA extraction from bacterial communities of different soils
E. V. Zaporozhenko, Natalia V. Slobodova, Eugenia Boulygina, Irina K. Kravchenko, Boris B. Kuznetsov
Abstract
E. V. Zaporozhenko, Natalia V. Slobodova, Eugenia Boulygina, Irina K. Kravchenko, Boris B. Kuznetsov
Abstract
A method for indirect DNA extraction from various soils significantly differing in their physicochemical properties has been developed. The proposed method is based on cell desorption from soil particles using a Tris-EDTA (TE) buffer supplemented with polyvinylpolypyrrolydone (PVPP) and sodium dodecylsulfate (SDS). Subsequent cell lysis and purification of DNA preparations methods based on alkaline lysis followed by chromatography on ion-exchange resins were described by us earlier. The purity of the DNA preparations obtained did not depend on the type of soil. It was shown that the DNA preparations can be used for the amplification of rather large fragments, e.g., sequences spanning the complete 16S rRNA gene.
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A method for indirect DNA extraction from various soils significantly differing in their physicochemical properties has been developed. The proposed method is based on cell desorption from soil particles using a Tris-EDTA (TE) buffer supplemented with polyvinylpolypyrrolydone (PVPP) and sodium dodecylsulfate (SDS). Subsequent cell lysis and purification of DNA preparations methods based on alkaline lysis followed by chromatography on ion-exchange resins were described by us earlier. The purity of the DNA preparations obtained did not depend on the type of soil. It was shown that the DNA preparations can be used for the amplification of rather large fragments, e.g., sequences spanning the complete 16S rRNA gene.
Key concepts: Lysis, DNA, DNA extraction, Extraction (chemistry), Alkaline lysis, Chromatography, Desorption, Chemistry