Wizard® Genomic DNA Purification Kit

DNA isolation / purification Bacteria - Gram positive Bacillus subtilis

Experiment
DNA isolation / purification Bacteria - Gram positive Bacillus subtilis
Product
Wizard® Genomic DNA Purification Kit from Promega
Manufacturer
Promega

Protocol tips

Protocol tips
- Incubate at 80°C for 5 minutes to lyse the cells; then cool to room temperature.

- Add 3µl of RNase Solution to the cell lysate. Invert the tube 2–5 times to mix.

- Incubate at 37°C for 15–60 minutes. Cool the sample to room temperature
Downstream tips
Rehydrate DNA by incubating 1 hour at 65°C, and then leave the sample at room temperature or 4°C overnight if DNA pellet difficult to dissolve

Publication protocol

Enzymatic DNA extraction with phenol/chloroform is a standard procedure in our laboratory21. Briefly, 10 µl of cell suspension of the respective strain in TE buffer was incubated twice for one hour at 37 °C after the additions of lysozyme and proteinase K, respectively. Following another 10 min incubation at 37 °C after adding sodium chloride and CTAB, the released DNA was separated from other cell components by the treatment with a combination of phenol and chloroform:isoamylalcohol in a 24:1 ratio. Finally, the DNA was precipitated using isopropanol, followed by a washing step with ethanol and the addition of 10 mM Tris pH 8.0 for resuspending the DNA pellet.

For the comparison of the extraction efficiencies with commercial kits, we used the QIAamp DNA Mini Kit from Qiagen and the Wizard Genomic DNA Purification Kit from Promega. The extraction procedures were carried out according to the manufacturers’ instructions for Gram-positive or Gram-negative bacteria, respectively.

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Discussion

Discussion

2 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

2 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

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Manufacturer protocol

Download the product protocol from Promega for Wizard® Genomic DNA Purification Kit below.

Download PDF Download manufacturer protocol

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