Ethanol substitute in Miniprep?

[jarlemag]

Hi. I've bought a miniprep kit which requires the addition of ethanol. Seeing as pure ethanol is difficult to get, I'm wondering if another alcohol, isopropanol for example, can be used as a substitute? 

Best regards,
Jarle

[Parijata D. Mackey]

Depending on where you live, it shouldn't be difficult (nor expensive)
to obtain: http://www.carolina.com/product/861261.do

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[Nathan McCorkle]

good link! what about USP grade (i.e. with nothing added)... that
would be ideal/desired for growth or enzyme related studies

Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

[ruphos]

On Sun, Aug 5, 2012 at 5:13 AM, Parijata D. Mackey <pari...@gmail.com> wrote:

Depending on where you live, it shouldn't be difficult (nor expensive)
to obtain: http://www.carolina.com/product/861261.do

It is worth noting that all the varieties offered by Carolina on that page are categorized as hazardous materials and only available to schools or businesses.

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-- Friedrich Nietzsche

[Nathan McCorkle]

On Sun, Aug 5, 2012 at 7:03 PM, ruphos <apokr...@gmail.com> wrote:
> On Sun, Aug 5, 2012 at 5:13 AM, Parijata D. Mackey <pari...@gmail.com>
> wrote:
>>
>> Depending on where you live, it shouldn't be difficult (nor expensive)
>> to obtain: http://www.carolina.com/product/861261.do
>
>
> It is worth noting that all the varieties offered by Carolina on that page
> are categorized as hazardous materials and only available to schools or
> businesses.
>

I read it as you need a commercial address... that means hackerspaces
and club rooms should be OK.

Using something like a Mailboxes etc account might work too (this
might only be in the U.S. though)
http://www.mbe.com/ps/Pages/ms.aspx

[jlund256]

If the prep requires 95% ehanol, you can use liquor store 190 proof alcohol, also called neutral grain spirits.  It is sold under the brand EverClear in the US.

 

If the prep requries 100% ethanol, you can often use 95%.  Adding 95% EtOH is adding 95% EtOH and 5% water, so if you are using it to precipitate DNA, add a bit more salt and a bit more 95% EtOH to get the desired final salt and EtOH concentrations.  This pushes up the total volume a bit.

 

Also, many protocols are not very sensitive, so adding 95% EtOH instead of 100% often will precipitate the DNA.  100% EtOH is a lab staple, so it gets specified even if the EtOH concentration is not a critical factor.

 

Jim

[Jeswin]

On Mon, Aug 6, 2012 at 11:19 AM, jlund256 <jlun...@gmail.com> wrote:
> Also, many protocols are not very sensitive, so adding 95% EtOH instead of
> 100% often will precipitate the DNA. 100% EtOH is a lab staple, so it gets
> specified even if the EtOH concentration is not a critical factor.
>
> Jim

The kits I use where "pure" Ethanol is needed say that 96% and above
can be used.

[Cathal Garvey]

You can't buy Ethanol that pure in Ireland or many parts of Europe. It's
literally illegal unless it's methylated, and in Ireland at least they
must also add azeotropic dyes and nausea-inducing chemicals to prevent
people drinking or distilling it.

So, given that you can't get technical-grade Ethanol without paying huge
duties and taxes over here, not to mention convincing D&E that you
should be allowed have Ethanol, I use Isopropanol almost all the time.

Isopropanol is actually better at precipitating DNA and protein,
requiring lower concentrations to achieve the same result. However, if
salt content is an issue, isopropanol-based DNA extractions may carry a
bit more salt than ethanol would.

Marked difference to watch out for; when precipitating DNA with ethanol,
the pellet is white and clearly visible (if it's large enough ofc). With
isopropanol, it's glassy and near invisible, so take care not to lose it.

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