Posted 26 May 2011 - 09:35 AM
So basically I made a glycerol stock and forgot to put it into the -80 C freezer. I realised this after 2 days at room temperature and put it immediately in the -80 C freezer. Would this affect the viability of cells, because being at RT will only increase their density.
Posted 27 May 2011 - 04:08 PM
Posted 28 May 2011 - 10:25 AM
Yes, it will decrease the viability. The cells will deplete the nutrients in the tube well before 48 hours are up.
Yes, but will I be able to use it as a glycerol stock to transform? If not, what are the chances of an unwanted mutation if I use 1 ul of my purified plasmid in the cell line and make fresh glycerol stocks ?
Posted 02 June 2011 - 01:17 PM
2 days at room temp is long.. most cells might be dead.
Mutations, I dont think there will be more mutations then when with regular growth.
Just make new cells... or are they so special?
Maybe you can just plate them out and make a new stock out of them? By doing this, you also immediatly see if they are still viable.. (plate out a few and if you see they are still viable, its worth a change using them. But to be honest: I would just use new cells, not worth losing a few days trying it with old cells.. but if you have some spare time you can try it)
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