DNA isolation / purification Bacteria - Gram positive Bacillus subtilis

DNA isolation and purification is the first critical step in sample preparation that helps ensure optimal performance of downstream assays like PCR, microarrays, and sequencing. Failure in yielding high-quality DNA would be the major reason that DNA doesn't work for the downstream application. To circumvent this, one should follow the recommended storage conditions to minimize DNA degradation by nucleases and shouldn't overload the purification system.

4 Matching solutions
2 Discussions

Found 2 discussions for this experiment

Discussion

5 years ago

5 years ago by Israel Lev 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?

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Discussion

5 years ago

5 years ago by 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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Found 4 matching solutions for this experiment

Wizard® Genomic DNA Purification Kit

Promega

Upstream tips	Protocol tips	Downstream 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	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

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

Manufacturer protocol Publication protocol 1 Relevant paper

DNeasy Blood & Tissue Kit

Qiagen

Upstream tips	Protocol tips	Downstream tips
		- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Downstream tips
- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Manufacturer protocol Publication protocol 1 Relevant paper

PowerSoil® DNA isolation

Mobio

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.

Manufacturer protocol Publication protocol 1 Relevant paper

Chromous Genomic DNA isolation kit

Chromous Biotech

Upstream tips	Protocol tips	Downstream tips
		- Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Downstream tips
- Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Manufacturer protocol Publication protocol 1 Relevant paper

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