HammerOfLight
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Electroporation help
HI,
I haven't been on much, being married really killed my spare time to work on my own projects. I have started up my microbiology lab again, although
on a smaller scale with a few equipment addition to what I used to use (the rest is in storage). I am starting from the beginning with culturing slime
molds, etc, working on pure cultures of fungi and bacteria, etc. I plan to do some DNA electrophoresis, and maybe some genetic transformation of
bacteria, and or plant tissue through electroportation. I've been reading on the subject, and have found a few things out, has anyone here done allot
of work in the field?
-Bad people are punished by societies law, good people are punished by Murphy\'s law.
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Reduce-Me
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I have, but I'll be damned if you can do electroporation at home.
Electrophoresis is another story. However, I'd stick with species that are genetically amenable to chemical transformations (unless you have the
tools)
You are going to end up cooking your cells to a sludge if you try to electroporate them in buffer that isn't deionized and/or if you can't control the
capacitance exactly.
Is there anything in particular you want to know?
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watson.fawkes
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Quote: Originally posted by Reduce-Me  | | I have, but I'll be damned if you can do electroporation at home. [...] You are going to end up cooking your cells to a sludge if you try to
electroporate them in buffer that isn't deionized and/or if you can't control the capacitance exactly. |
The
electronics problems seems completely solvable, even at the amateur level. There's no real problem timing microsecond-resolution pulses with today's
microcontrollers and power switching devices. A switching network can give you a fair measure of programmable capacitance. Voltage regulation is easy.
Are there any other pulse parameters that are relevant, such as the shape of the discharge pulse?
[Grammar edit.]
[Edited on 15-3-2010 by watson.fawkes]
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Reduce-Me
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yes, there is, and that (shape, voltage and resistance) is exclusively dependent on species.
Even though the electronics issue might be solved, there is still the issue of a cuvette to keep the cells in during the shock. Dimensions can be
looked up. But the cuvette they are held in is just as important, because the specifications of the cuvette dictate the type of pulse(s) you would
need. The type of metal to transfer the shock the distance between the plates etc.
Relevant options are...
Capacitance
Voltage
Resistance
cuvette type.
the time the pulse takes is dependent on the cuvette more than anything.
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HammerOfLight
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I actually was referring to
I was referring to actual electroporation equipment and curvettes, I have found equipment on e-bay from time to time. I also want to put together an
Electrophoresis apparatus as well for some DNA work. I was looking at the ones in NASCO's website (www.enasco.com).
-Bad people are punished by societies law, good people are punished by Murphy\'s law.
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Reduce-Me
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http://www.pearlbiotech.com/diy
has built in transilluminator.
You can rig some sort of power supply.
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