That'll be one of my next projects... :) An electroporator....
Using an piezoelectric lighter.... Has anyone a foto of the circuit?
You have to push the piezoelectric botton with your hands very often,
or is there a circuit that transforms the voltage??
On 3 Feb., 19:55, Laureana Stelmastchuk <laure.stelmastc...@gmail.com>
wrote:
> Well, it wont answer your main questions, but could give you some great
> ideas...
> Have you ever tried electroporation? (http://en.wikipedia.org/wiki/Electroporation#Electroporators).
> I think electrocompetent cells are easier to prepare than the chemical
> ones, and it shows better transformaton rates. I have used both *E.coli *DH10B
> and DH5α.
> However, if you don't have an electroporator... I don't know, but would be
> cool to try to make one.
>
> 2012/2/3 Jonathan Nesser <jonathan.nes...@gmail.com>
>
>
>
>
>
>
>
>
>
> > I posting this as another question in the form of "I must be crazy, please
> > correct me." My understanding of induced competence in e. coli is that it
> > involves creating a new culture in LB medium, soaking in a CaCl solution
> > (other components can be added for the perfectionist), cooling and then
> > heat shocking, as well as being very gentle with the cells (no vortexing,
> > etc). I know that many companies sell e. coli cultures that have been
> > deemed competent, but is there any reason why using the above procedure on
> > new cultures from other sources which are incompetent would not yield
> > competent cells?
>
> > My problem with the standard lab method of simply ordering competent cells
> > is (besides the obvious expense factor), that I don't have any means of
> > storing the competent cells I receive at -80C, which I'm told destroys the
> > company's competence work anyways. Also, I've discovered that I can obtain
> > cultures for the Coli Genetic Stock Center at Yale for substantially less
> > (less than half that which New England Biolabs charges, and the price for
> > extra strains in the same order increases te savings exponentially). It
> > seems like I must be missing something somewhere, because it doesn't make
> > sense that large labs wouldn't induce competence themselves (unless it's
> > just a time saving and quality assurance factor).
>
> > Also note that I have read that most "home" induced competence jobs yield
> > a lower transformation rate, but figure that with selection and time to
> > grow cultures I should be able to yield a decent number of transformed
> > cells in the end, and in the case of everything I will be doing in the next
> > half year, there is no product I'm looking to yield (unless I get into
> > cubcloning), just the confirmation that my transformed plasmid works.
>
> > Someone please set me straight, and as always, I apologize for the simple
> > question, and deeply appreciate the time spent answering my questions.
>
> > Jonathan Nesser
> > jonathan.nes...@gmail.com
> > diybioandneurosci.blogspot.com
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> --
> Laureana Stelmastchuk Benassi Fontolan
> Lab. Biologia Molecular de Coccídias
> Depto. Parasitologia
> ICB II - USP
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