Hello everyone, was just wondering if it is normal for transformed E. coli to become pelleted in a falcon tube when left for overnight culture in shaky incubator at 150rpm ? That is because i think normally bacteria are properly suspended in media when left in shaky incubator. Any thoughts would be appreciated. Thanks
Thank you for your reply. Yes, that is what we do, by plating on LA plates with antibiotics and it is understood to reach to stationary phase, but in my previous work with bacteria, the bacteria is usually well suspended in broth while left overnight in shaky incubator where as my transformed E. coli usually pellets down in tube after overnight in shaky incubator. I am just wondering is this normal and fine ? Many thanks in advance
It could be... they aggregate, possibly because of the stress or because you have transformed it with something that favours aggregation...
I have worked with some wt strains that sometimes produced clumps and then deposited at the bottom of the flask.
I think if everthing works fine you don't have to worry, but you can also search the cause. It could be interesting...
Your cloned insert may be inhibiting cell division. I have found this when cloning adhesion molecules and "sticky" proteins. Shaking at 250 RPM may keep your cells in suspension.. Finally, try securing the tube in the rack, it should improve the culture mixing. Or, use a small Erlenmeyer flask.
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