one or two sparks should be sufficient as long as the volts/cm is right.
On Tue, Feb 7, 2012 at 8:14 AM, Mega <masterstorm123@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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