PowerSoil® DNA isolation

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Do not use bleach to clean the inside of the PowerVac™ Manifold or to rinse the PowerVac™ Mini Spin Filter Adapters when attached to the manifold.

Upstream tips
Do not use bleach to clean the inside of the PowerVac™ Manifold or to rinse the PowerVac™ Mini Spin Filter Adapters when attached to the manifold.

Publication protocol

DNA extraction
The PowerSoil DNA Total Isolation Kit (MOBIO Laboratories Inc. Solana Beach CA, USA) was used for genomic DNA isolation and the manufacturer’s protocol was optimized for the samples as follows: after the bead beating lysis procedure and three cleaning solution steps, TE buffer was added to the C3 solution to its final concentration (10 mM Tris, 1 mM EDTA, pH 8.0) and samples were incubated at 55 °C for 1 hour with proteinase-K (Sigma-Aldrich, 100 ug/ul).

Two Phenol:Chloroform:Isoamyl alcohol (Sigma-Aldrich) and one chloroform purification steps were carried out, followed by an ethanol precipitation with ammonium acetate (2 M final concentration) at −20 °C. Samples were washed twice with 75% ethanol and after drying, pellets resuspended in 100 μl of molecular biology-grade sterile DNAse-RNase free water (Sigma-Aldrich).

Purity and quality of each extracted DNA were measured by NanoDrop 1000 spectrophotometer (Thermo Scientific) and Qubit Fluorometer (Life Technologies) DNA quantification systems. Three extractions were performed for each sample and nucleic acids were finally pooled together.

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Papers

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Manufacturer protocol

Download the product protocol from Mobio for PowerSoil® DNA isolation below.

Download manufacturer protocol

Videos

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