I have a mixture of bacterial and plant cells which is yielded from two different methods:
1) vigorous shaking of plant tissue which have bacteria living on it.
2) the same plant tissue is blended in liquid nitrogen.
I want to make sure if centrifuging these two will separate bacterial cells from plant cells and does this process mechanically lyse either bacterial or plant cells.
How would you be able to separate the cells by centrifuge ? Are you talking about normal centrifuge or density gradient centrifuge.
I suppose you would not be able to separate them. And centrifuge is no way to lyse either bacterial or plant cells.
Please describe more about the material or experiment you have. Why do you want to separate that?
If you want to have pure cell collection/culture, you have to consider isolating them separately. You have to surface sterilize the plant material, to make sure there is no bacterial contamination. To have pure bacterial culture, you have to grow the plant surface microflora on culture media (culturable microflora). Them you can proceed to use them separately for whatever your purpose is. But as i said, explain more about your problem to get better answers.
@Sourav - can you please explain Moghimi how you justify your answer?
Dear Abhijeet Singh;
thank you for your time and answer. My goal to separate bacteria from plant cells to perform an NGS analysis on bacteria population. So, the bacterial population is important. But why I do not perform culturing them on culture media to purify bacteria? The answer is that many of anaerobic bacteria will lose their viability even in short exposures to air. I have a defined protocol from a paper published in a high rank journal. 2017 and I am investigating all your questions through my question. That if the centrifuge alone can separate bacteria from plant cells?
Now, if Sourav Maiti is right and bacteria don't get lysed I think the problem is solved. Cause after centrifuging the mixture there will be bacterial cells and so many cell organells (from plant cells) and macro-molecules which makes bacteria to form pellets and all others are supernatant and can be removed. If not, there is the problem. As the researchers who published the paper I mentioned have reached reasonable results, I assume they have done it right. But still I want to make sure.
I will appreciate anything else that comes to your mind.
What analysis of bacterial populations? If it is 16S rDNA sequencing, you can use bacterial universal primers. You dont have to separate them. One thing to mention, you don't have to care about if the bacteria is dead or alive as you are not doing transcriptome analysis.
Yes it is 16s rDNA sequencing, and yes I understand what you mean but still the problem is that we need a pool of DNA with the minimum amount of plant DNA, cause then we will ask some lab to analyse a certain amount of extracted DNA and if there is too many of plant DNA then the chance of small populations of bacteria to be revealed will get very low, instead plan DNA are shown which will be omitted.
The other point is that I don't care if they can live or die, but culturing them in a medium that is favorable for some and unfavorable for others will change proportions of different bacteria from the actual profile.
Aren't I right?
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