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Pellet Formation


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#1 Faith

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Posted 07 July 2010 - 11:29 AM

Hi there,

Are there certain concentrations and volumes at which e.coli become visible as a pellet after centrifugation?

I am trying to stain e.coli with acridine orange and would like to remove the "fluorescent background" of the stained liquid I have the cells suspended in (which at the moment is PBS). I would like to replace the stained PBS and introduce fresh PBS so that only the cells are stained.

I am trying to do this for concentrations of about 10^6 and 10^7 bugs/mL. I can get a pellet when I spin the original amount (~3*10^8) in a 5mL volume.

Any suggestions?? Thanks!!

#2 perneseblue

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Posted 19 July 2010 - 06:46 PM

you could try spining harder but for a shorter period of time. Try 14000rpm for 30 sec
May your PCR products be long, your protocols short and your boss on holiday




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