If you have a plate culture, pick a colony and add to 10 ul of sterile TE buffer. Heat at 95 degrees celsius for 10 minutes. Centrifuge and use the supernantant for downstream processes.
Please go through the link
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2710730/
If you have a plate culture, pick a colony and add to 10 ul of sterile TE buffer. Heat at 95 degrees celsius for 10 minutes. Centrifuge and use the supernantant for downstream processes.
Even simpler than that, if you are doing PCR from a culture, you can just take a scrape of a colony with a pipette tip and wipe it into the bottom of the PCR tube. Add the PCR reagents on top and run the PCR. The initial 95deg step is enough to break down the cells. Of course it all depends on what you are trying to achieve. This method is only good for imprecise qualitative methods.
It depends on the source of the sample you want to analyze.
If pure culture of bacteria: Boiled 100 ° C for 10 minutes.
If your source is from microbiota, I recommend you look column-based kits. Depending on the source of the microbiota (soil, plant material, water etc) some kits work better than others.
David Ian Walker and Félix Morán have it right. I would just say make the initial heating step for the pcr 5-10 min rather than the standard 2 min at 94˚ or 95˚ C for pure culture or colonies on a plate. Then second easiest would be to purchase a kit, either through Mo Bio or Zymo. For low molecular weight and or high amounts of inhibitors there are numerous more complex ways you can go.
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