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glycerol stock preparation of recombinant clone


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#1 biotef

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Posted 19 December 2010 - 09:42 PM

we have recombinant e.coli clone used for preparation of cytokine. we preserve this clone in glycerol stock at -80 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

#2 protolder

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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 phage434

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Posted 20 December 2010 - 05:05 AM

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




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