[DIYbio] Re: competence in e. coli

You don't need any resistors or capacitors or ... anything?

Just an ignitor, electrodes and a cuvette??????

If so, Awsome!!!

On 7 Feb., 17:49, Nathan McCorkle <nmz...@gmail.com> wrote:
> one or two sparks should be sufficient as long as the volts/cm is right.
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> On Tue, Feb 7, 2012 at 8:14 AM, Mega <masterstorm...@gmail.com> wrote:
> > 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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>
> --
> Nathan McCorkle
> Rochester Institute of Technology
> College of Science, Biotechnology/Bioinformatics

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