Re: Get a plasmide - buy? share?

In our university's lab there are big ammounts of 'pure' ethanol
(97%). I think isopropanol is more difficult to obtain for me.

On 26 Dez., 19:49, Cathal Garvey <cathalgar...@gmail.com> wrote:
> 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 !!
>
> --www.indiebiotech.com
> twitter.com/onetruecathal
> joindiaspora.com/u/cathalgarvey
> PGP Public Key:http://bit.ly/CathalGKey

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