we have recombinant e.coli clone used for preparation of cytokine. we preserve this clone in glycerol stock at -80º C. we prepare 50% glycerol stock autoclave it and mix log phase culture to it to get final glycerol conc of 10%. Kinldy let me know if procedure is O.K as one of my seniors mention that i should centrifuge the culture suspension wash it and then suspend the pellet in glycerol to get 10% concentration. Which technique is better. kindly advise.
thanks in anticipation
biotef
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#2
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Posted 19 December 2010 - 10:39 PM
Hola, I always mix 0.8 ml of culture or resuspended pellet with 0.2 ml of sterile concentrated glycerol (87%). The advantaje of freeze the resuspended pellet is that when you want to start a new culture your inoculum will be stronger with more possibilities of grow. Remember that the seed is made of the frozen stock without thaw it. Buena suerte
#3
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Posted 20 December 2010 - 05:05 AM
Your approach is exactly what I do. It has always worked.
