Electrocompetent cells

D.K. dk at no.email.thankstospam.net
Tue Oct 22 22:48:56 EST 2002


 Philipp Wechner <philipp.wechner at uibk.ac.at> wrote:
>Same protocol here. Use Cells from a culture between OD600= 0.5-0.8. After
>the culture reachted 0.8 the cells are getting worse for electroporation.

All these numbers in all protocols are of dubuous value because 
OD600 is spectrophotometer-dependent reading. What spec did
you use? 

DK

>"Bryan J. Maloney" schrieb:
>
>> Something that gave success fairly routinely was growth to late log phase
>> in YENB (0.8% Yeast Extract, 0.8% Nutrient Broth, nothing else).  Then
>> wash twice in cold water, once in cold 10% glycerol, and resuspend in cold
>> 10% glycerol.  Then we froze the cells on powdered dry ice/ethanol, pre-
>> chilling the tubes.  One trick was that the cells were remarkably fragile,
>> so we had to spin them very gently when washing.
>



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