PowerSoil® DNA isolation

DNA isolation / purification Bacteria - Gram positive Enterococcus faecium

Experiment
DNA isolation / purification Bacteria - Gram positive Enterococcus faecium
Product
PowerSoil® DNA isolation from Mobio
Manufacturer
Mobio

Protocol tips

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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Discussion

Discussion

5 years ago

Author: Israel

How can I improve my DNA yield?

The DNA concentration after using this DNA isolation kit is sometimes too low and thus it is not sufficient for my follow-up experiments. How can I improve it?

Discussion

5 years ago

Author: Milena Alexeyeva Russian Federation

Tips on storing DNA templates?

Hello there! I just started doing experiments on bacterial DNA and I would like your opinion on storing DNA templates. Which are the desired and most optimal conditions?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA isolation / purification Bacteria - Gram positive Enterococcus faecium using PowerSoil® DNA isolation from Mobio.

Paper title
Direct 16S rRNA-seq from bacterial communities: a PCR-independent approach to simultaneously assess microbial diversity and functional activity potential of each taxon
Full paper
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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