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Resuspending Cells

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Resuspending Cells

Postby NiceOnTheIce » Wed Nov 17, 2010 5:38 pm

For my lab tomorrow there's a step that says:

"Add ~2mL of ice cold 0.1M CaCl2 to the cell pellet and gently resuspend the cells." I know what resuspending means but I don't know how to go about doing this, as we are supposed to pour the CaCl2 straight into the tube (approx ~2mL) and can't stick any instrument in there, to prevent contamination. Do I shake it up?

Thanks,

Holly
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Postby protold » Thu Nov 18, 2010 6:24 am

Hola, It is supossed that you work in a culture hood with all the material inside sterile, so you add 2ml with an automatic pipette, or plastic sterile pipette or sterile glass pasteur pipette and sucking up and expeling liquid the cells resuspend with the moving. when the suspension is homogeneus you take it and seed a new plate. Good luck
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Postby JackBean » Thu Nov 18, 2010 6:44 am

as protold said, you are supposed to work in sterile environment, otherwise you would contaminate your own sample. So you can simply pipette the solution with pipette up and down. Or you could use vortex.
However, you should work carefully, otherwise you could damage (kill) your cells!
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Postby NiceOnTheIce » Thu Nov 18, 2010 12:40 pm

That makes sense, but we were specifically told that we would not be using glass pipettes (just estimating the amount of CaCl2 according to the lines on our centrifuge tube) and that vortexing would kill our cells. So I still don't know what to do.
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Re: Resuspending Cells

Postby protold » Mon Nov 22, 2010 6:51 am

Hola, as Jack says, pipeting carefully up and down with an automatic microppipette (as Gilson) with a sterile plastic tip, or a 2, 5 ml plastic sterile pipette or a sterile plastic squizzer you can solve your problem. Cheers
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