I want bacterial cells apart from plant cells. I have seen some papers do that exactly as I need (separating bacteria from cereal silages) but as they don't explain it that much, I am a little confused.
can anybody help? this is what I have found:
" silage samples were added to a 20× volume of sterilized phosphate-buffered saline (pH 7.4), and extraction was performed by vigorous shaking for 10 min at ambient temperature. Bacterial pellets were obtained by centrifugation at 8000×g for 15 min, then were washed once with 1 mL of solution containing 0.05 M D-glucose, 0.025 M TrisHCl (pH 8.0), and 0.01 M sodium EDTA (pH 8.0). After centrifugation at 15,000×g for 2 min, bacterial cells were lysed with 180 μL of lysozyme solution (20 g/L lysozyme, 0.02 M Tris-HCl [pH 8.0], 0.002 M sodium EDTA [pH 8.0], 1.2 g/L Triton X-100) at 37 °C for 1 h. "
Try the techniques in the following links
http://aem.asm.org/content/83/5/e03151-16.full
Article A Novel Technique for Feeding and Confirming Uptake of Bacte...
Instead of phosphate buffer you can use Nutrient broth or other growth media that allow all bacterial cells to grow/ enrich.
Dear Bhupender Sharma ; Abdallah Hussien Fathy ;
I'm afraid this leads to change in ratios of bacteria and as I am planning to assess them qualitatively and quantitatively, it will be a source of bias.
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