A lab-made method for extracting DNA from soils

• DOI: 10.1071/sr17261
• Publication Date: 2018-07-17T00:27:40Z
• Authors: Michael Anderson
• Keywords: 0301 basic medicine; 03 medical and health sciences

Most soil biologists rely on proprietary commercial kits to extract DNA for sequencing projects, primarily their ease of use. However, most these lack procedural transparency which limits comparative long-term standardisation efforts. Here I demonstrate a new procedure that is well defined and yields results similar or superior the PowerSoil SoilMaster kits. The Laboratory Technique utilises bead-beating, well-defined extraction buffer, ammonium acetate precipitation, isopropyl alcohol precipitation purification through Sephacryl 300-S HR gel filtration spin column. Based Sybr Green assays extracted average 21% 430% more than respectively. Purity based 260/280 nm spectrophotometric ratios averaged 1.94 procedure, 1.35 kit. Correlation between community taxonomic structure analysis from by were in near complete agreement yielding an R2 0.99 at level phyla 0.97 genera. Overall, attractive alternative terms yield, purity, transparency, cost potential standardisation.