On Sat, Mar 10, 2012 at 12:56 PM, Mega <masterstorm123@gmail.com> wrote:
> It would surely be worth a try!
> 25kV/cm for E.coli?
>
yes that's what works for most people, but a little higher or lower
should be fine (its just an efficiency thing, but electroporation is
way more efficient than CaCl2), I think the sparkers put out ~18kV
> But isn't it better if I have a successful standard-transformation
> done before trying something more difficult??
>
You've tried once or twice already without it working... the
instructions I just gave are
pretty simple, I think its worth a try! (Just make sure your amp
concentration is right this time!)
>
> On 10 Mrz., 15:00, Nathan McCorkle <nmz...@gmail.com> wrote:
>> Mega,
>> you should just try to add E.coli from a plate to sterile water in a
>> microtube. Add your DNA, then place the electrodes from a piezo
>> sparker in and spark 1-3 times (or have 3 tubes, and try 1 spark on
>> tube #1, 2 sparks on tube #2, 3 sparks on tube #3). Then add some LB,
>> and incubate for 1 hr... then plate that on LB + amp plates.
>>
>> No CaCl2 or anything... its an experiment, but you could be the first
>> to actually try it! (we've talked about it a lot on here, but never
>> tried it)
>>
>>
>>
>>
>>
>>
>>
>>
>>
>> On Sat, Mar 10, 2012 at 8:19 AM, Mega <masterstorm...@gmail.com> wrote:
>> > @
>> > Chris:
>>
>> > That's amazing!!!
>>
>> > Did I understand that correctly? Usually you transform roughly 1
>> > cell and 999 stay untransformed. Using this technique, ~600 out of
>> > 1000 cells are transformed???
>>
>> > If that is true, I must ask a friend of mine (electrical engineer)
>> > if he is interested to build such a miracle device!!!!!
>>
>> > --
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>>
>> --
>> Nathan McCorkle
>> Rochester Institute of Technology
>> College of Science, Biotechnology/Bioinformatics
>
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--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics
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