On Sat, Mar 10, 2012 at 2:44 PM, Jeswin <phillyj101@gmail.com> wrote:
> On Sat, Mar 10, 2012 at 2:32 PM, Nathan McCorkle <nmz787@gmail.com> wrote:
>
>>>
>>> What do you mean by rinsing with water?
>>
>> From my LB liquid culture I centrifuged, decanted supernatant,
>> resuspended in distilled/sterile/nanopure water, repeat once or twice
>>
>
> So these cells are the regular, hardy kind? The competent cells
> normally used are treated with utmost care, like no rapid temperature
> changes, and they lose transformation efficiency per freeze/thaw
> cycle.
The water rinse and electric shock was all I did to 'make them
competent', straight from an 3-6 hour culture which was a subculture
of an overnight culture. For competency to really mean anything, it
needs to be associated with a transformation efficiency number...
doing this gave me higher efficiency than CaCl2 transformation, but
certainly lower than if I would have taken the time to 'prepare
electrocompetent cells', being more careful with the temperatures,
etc...
>
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--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics
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