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What exactly does it mean to "resuspend the cell pellet"?

Asked by ScottyMcGeester (927 points ) March 24th, 2014

I can’t go into much details about the experiment, but I’m not getting any results from trying to obtain DNA from E. coli cells.

I have E.coli cells in a pellet in a tube. The very first step is to resuspend it using this particular buffer. I know the problem I’m having has to be at the beginning because the setup is in such a way that I can tell where the issue must be if I get absolutely no results from controls or the test samples.

(If it isn’t the resuspension part, then my E.coli cultures must be bad, but I doubt it because they’re fresh and have a high absorbance)

When I do that resuspension step, I pipette the required amount into the tube. . .but the pellet doesn’t necessarily “suspend” in the liquid. It just stays there at the bottom. I invert the tube a little and part of the pellet breaks off. I’ve been told to always “be careful” with the pellet, so I’m confused as to what “resuspension” then should LOOK like when using it to resuspend a cell pellet. Do I have to break up the cell pellet?

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