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glycerol stocks - (Mar/07/2008 )

What % glycerol should be used for DH5 alpha cells? I find lots of variation in what people are using for glycerol stocks....

-katjo-

We just did the experiment. You can grow cells from 7.5 to 15% glycerol in LB. Less than 7.5% will not cryoprotect cells. More than 15% becomes inhibitive to growth. We grow cells in LB + 10% glycerol. If you add glycerol after cells are grown, then much more glycerol can be added, probably up to about 50%, but we didn't check that.

-phage434-

QUOTE (phage434 @ Mar 8 2008, 05:58 AM)
We just did the experiment. You can grow cells from 7.5 to 15% glycerol in LB. Less than 7.5% will not cryoprotect cells. More than 15% becomes inhibitive to growth. We grow cells in LB + 10% glycerol. If you add glycerol after cells are grown, then much more glycerol can be added, probably up to about 50%, but we didn't check that.

15% glycerol or 7% DMSO are tthe optimal amount for cyroprotection of bacteria.

-party-

To prepeare a stock, i grow your DH5 cells in LB overnight and add glicerol to final concentration of 25%.
i make all my stocks like this and I have no problem at all.... blush.gif

-solmaniar-