glycerol stock preparation of recombinant clone - (Dec/19/2010 )

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

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

Your approach is exactly what I do. It has always worked.