Re: Get a plasmide - buy? share?

Well, "Supernatant" = liquid layer, so the supernatant is what you want
to collect for plasmids. The pellet, meanwhile, is all the stuff that's
formed from precipitation of cellular matter.

To isolate the DNA, you will also need alcohol and perhaps sodium
acetate. For alcohol, you can either use ethanol (if you can get pure
ethanol where you live) or isopropanol (available anywhere). You need
different amounts of whichever one you use, and the precipitated DNA
will look a little different (white with ethanol, glass-like with
Isopropanol), but the DNA will be fine either way.

If you need sodium acetate (I can't remember, check the protocol), you
can either make it by reacting acetic acid and sodium carbonate (vinegar
and baking soda), or more preferably buy it online from a pure chemicals
website such as mistralie.co.uk.

Alternatively, those re-usable hand warmers that contain a little metal
tag you bend to activate them? Which you boil to re-melt for reuse? That
liquid is sodium acetate trihydrate, so you can use it provided you
calculate the amounts including the extra water (you'll find a
comparison of the molar weights on wikipedia), and you should use it as
a *solid*, not a liquid. The liquid form may spontaneously crystallise
(I think?) on contact with other stuff like leftover cell matter,
releasing tons of heat and inconveniencing your experiment. :)

On 26/12/11 16:07, Mega wrote:
> As soon as I get the lab strain, I'll do a boiling lysis...
>
> That seems quite easier than an alkaline lysis, because the only
> chemical substance I need is STET-buffer. And this one substance I'll
> get anyhow.
>
>
> The supernatant 'll be junk and within the liquid there will be the
> plasmids (and some proteins that won't affect further transformations
> very much) then?
>
>
> ____
> Merry Christmas @ all !!
>

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