Washing cells with...? - (Oct/15/2008 )
When it comes to harvesting and washing bacteria cells, how fussy are you during the washing part i.e. do you use water or buffer? I have to harvest my cells, wash them and then transfer them into liquid medium to grow for another period of time. Plan to use PBS but if plain ddH2O can be used then I shan't prepare them. One suggested 50mM KH2PO4 but then I find no reports of using that conc of KH2PO4 which will prevent changes to the cells.
PBS ...not ddH2O
Agree with Minnie....... PBS too
If it's just to set up a new culture you can also wash with you medium.
For most bacteria, it makes little difference as long as you don't leave it a long time. What do you plan to do with the washed cells?
Plan to transfer them for culture in a limited/minimal medium. Don't want to have carry over of nutrients from 1st medium. Some used distilled but I'm not sure if it'd lyse the cells. If not, then I'd opt for the easier ones.
Centrifuge 12K at 2 mins / 4'C. Alternatively 8K 10 mins
Just washing with sterile water will not bother most bacteria, assuming final destination is a more osmotically-friendly environment. Use physioloigcal sailne if you're worried - I'd not bother with the buffer.
Thanks George. I'll go with the PBS. Thanks again everyone.