competent cells again!!

gwallmanning at my-dejanews.com gwallmanning at my-dejanews.com
Wed Jun 3 18:58:50 EST 1998


In article <l03130300b19b60eeb7ca@[134.174.248.196]>,
  jaugliera at RICS.BWH.HARVARD.EDU ("John Augliera") wrote:
cold CaCl2 and placing at -70C.  It makes no mention of adding a
> cryoprotectant such as glycerol.  Would anyone know if this is OK!! or
> would the addition
> 10% glycerol adversely affect the competency or viability of the cells.
>
> Thanks
>
> John Augliera
> Technician
> Brigham and Women's Hospital
> Boston,MA 02115

Adding 10% glycerol does not appear to affect the competency of the cells,
although I would think that it would enhance the viability.  You could split
a batch and treat half with and half without glycerol, and test competency
and viability.  In the past I have found that it is better to filter
sterilize the glycerol solutions than autoclave them as the autoclaving
apparently turns enough of the glycerol into aldehydes etc for it to be toxic
- maybe a chemist will correct me I don't know, but I couldn't get bugs to
grow on autoclaved media containing glycerol, but they would grow fine if the
glycerol was filtered.

Good luck

Glenn

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