Technique / Proteomics and protein biochemistry / Protein expression, protein extraction and protein purification


Preparation of Glycerol Stocks and Thawing of E.Coli Stocks



Preparation of Glycerol Stocks and Thawing of E.Coli Stocks

The BL21 cells are fairly fussy in that, unlike other E. coli strains, their plasmids become unstable if the cells are stored in glycerol concentrations greater than 10%. For this reason, the cells should be stored in 8% glycerol and frozen quickly in liquid nitrogen. (Structural biology Protein preparation protocols, South Africa Structural Biology Initiative)

Last update 30-Dec-2005, Rating of 1 votes.

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By Peter Kim on 30-Dec-2005
This protocol does not make sense- there is an error either in the description or the protocol.

It starts off by saying that the plasmids are unstable at glycerol conc >10% so the cells will be stored in 8%. Good so far.

The protocol then calls for mixing 100ul of cells with 900ul of an 80% glycerol mixture. That would give a final concentration of 72%.

I wonder if that should read 100ul of 80% glycerol with 900ul of cells. That would give a 8% final conc.
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