Pelleting bacterial cells - (Mar/18/2013 )
I am trying to do enrichments from some seawater that I have. I would like to pellet down cells from my triplicates and add them to my media. I was going to centrifuge about 50mL at 3000g. Does this sound right to anyone? I don't want to hurt the cells so it needs to be gentle. I also was wondering if I should do 50mL of each of my triplicates then add the three pellets or a combined 50mL of the water. How can I get the pellet out without hurting the cells? Could I resuspend the pellet in the media and add it to the remaining media? THanks for any help
Hurting bacterial cells? You probably will damage them more with changing the media that with centrifugation, they aren't mammalian cells or whatsoever. Most bugs even are able to grow at 100000g. Hope you have a lot of cells in your sample because at 3000g will take a while...
All depends how much of the biomass you want to recover, if you want to recover all the cells or at least minimise the bias during the recovery,... cells with different shapes and sizes take different times to pellet. Check this for example:
20000g x 20 min gives a quite even recovery