Nitrous oxide cavitation
Meredith L. Patterson
That is a pretty damn clever way to do a nitrogen bomb.
--mlp
ByoWired
On Dec 2, 8:57 pm, "Meredith L. Patterson" <clonea...@gmail.com>
wrote:
> That is a pretty damn clever way to do a nitrogen bomb.
decompression, those types of devices routinely utilize 2000 psi or
more, which is far far higher than this whippet device. If you were
crazy enough to try to pump 2000 psi into that thin-walled whippet
container, the only cells you would end up lysing would be those of
your own.
Patrik
water, which is why you need the high pressure. N2O dissolves about
seven times better, so you should be able to get away with a much
lower pressure.
One problem may be that N2O might act as a catalyst for oxidation -
that's what it seems to do in race engines anyway. Either way, it's
worth a try. If your $30 whipper doesn't work as a cavitation vessel,
you can always make some awesome cocktails with it...
Patrik
Jonathan Street
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Simon Quellen Field
Patrik
>
> If the gas dissolved in lipid bilayer membranes and then came out of
> solution suddenly, cells walls might be affected.
whether a N2O whipper can used to lyse cells, in place of nitrogen
cavitation. If N2O specifically targets cell membranes, this could
indeed be a good solution.
Cathal Garvey
About time we found out. Anyone with a small cream whipper, yogurt and a centrifuge handy? Protocol I'm imagining:
Spin down gram positive yogurt bacteria, wash in water to get a clear pellet of cells.
Suspend in a small volume of water or TC/TE, put in whipper, charge whipper with N2O, shake.
Discharge whipper in vertical orientation as quickly as possible (wear gloves in case of freezing), to decompress cells. You could push cell suspension out like cream but this may shear DNA?
Quickly check your suspension for disruption: gloopy mess is disrupted cells? Don't know what to expect here, I'm used to alkaline lysis minipreps.
If you used tris-EDTA or tris-citrate* to resuspend the DNA might survive long enough to check: take half to gel run as-is. Hope for a slow-moving band of genomic DNA complexed with protein.
Spin remaining suspension at 10k RCF to clarify, take supernatant and run in agar (or agarose if handy) gel immediately, stain with whatever's available, probably methylene blue, hope you see plasmids alone.
It's possible you'll need better cleanup even if it works before you'll see any DNA result, but even a strawberry-style extraction (salt and alcohol) on the disrupted cells will remove a lot of protein and fat crud and improve chances of resolving DNA.
*as yet untested as a creating buffer for DNA. Replacement of tris with another alkaline buffer (but not calcium/magnesium salts, please) might work equally well, for quick and dirty preps. Totally unknown here though.
Sent from my Android.
On 6 Dec 2010 07:44, "Patrik" <pat...@gmail.com> wrote:
On Dec 5, 9:39 am, Simon Quellen Field <sfi...@scitoys.com> wrote:
>
> If the gas dissolved in lipid...
And that would be a good thing, right? After all, we're discussing
whether a N2O whipper can used to lyse cells, in place of nitrogen
cavitation. If N2O specifically targets cell membranes, this could
indeed be a good solution.
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Jay Woods
It would be interesting if a pop bottle soda siphon would work with the
N2O cartridges.
It would be interesting if a CO2 cartridge would work with the Cream
Whipper.
It would be interesting if there were a head available that would work
with a pop bottle and an N2O cartridge instead of having to buy a
stainless steel bottle/head or a whole head/stand.
Patrik
> A couple of thoughts. [...]
From reading the various threads that spun off the original "Infusion
Profusion" article, I believe the answer to all those questions is
yes. This is mostly just based on what I remember, so grains of salt
notwithstanding:
- a soda siphon should work, but tends to operate at lower pressure,
and may not be able to release the pressure as fast (i.e. less
cavitation)
- CO2 cartridges do work on the whipper as well, but you wind up with
a carbonated liquid, i.e. pH change due to carbonic acid, and lots of
residual dissolved CO2 at room pressure. In contrast, the N2O pretty
much all comes out of solution at room pressure.
- In the Comments section of the original article, "Mr. Fizz"
mentioned alternatives to using a whipper:
"Use plastic jugs when you can get them. For example, Coors & Miller
Lite are both sold in little kegs now. They’re known as Beer In A Box.
Drink the beer. Save the kegs. There’s already a 16-gram co2 cartridge
dispenser attached. The size of the mouth of the keg is larger than
that of 2-liter soda bottle, so you can get larger food item in it.
You can even get Fizz Giz caps to fit these kegs. With them, you can
introduce pressure using any gas you wanna buy, o2, air, n2o, co2,
helium, nitrogen, acetylene, propane (just joking on a few of those).
Check out my Fizz Giz website and google for pressure infusion
marination."