May have incorrectly thawed a bacterial culture from -80 C
Posted 28 June 2012 - 01:53 PM
So I'm trying to create a fresh LB culture (in order to later maxiprep) from a stock frozen at -80. Initially I had the culture thawing at room temperate for maybe 3 to 4 minutes (which I no know to be wrong) before I placed it back into the -80. I put it back so I can first figure out what the hell I should do in order to not kill the bacterial stock (if I haven't already).
So now my PI has told me the best way to do this is to pull the culture out stick a torched inoculating loop in there, scrap up some gunk and quickly transfer to the warmed LB broth. Then throw back into -80 immediately.
In your guys' experience, is that 3 to 4 minute initial room temp period enough time to destroy a stock like this? For the most part I saw that the culture still remained solid as I was placing it back to -80.
Posted 28 June 2012 - 02:01 PM
Edited by bob1, 28 June 2012 - 02:02 PM.
Posted 28 June 2012 - 02:47 PM
Posted 29 June 2012 - 06:11 AM