mini-preps: NOT kits.

Michael Szardenings msz at bio.embnet.se
Sat Sep 23 08:16:08 EST 1995


In article <1995Sep21.125045 at ania.path.ox.ac.uk>, rpgrant at molbiol.ox.ac.uk
(Mad Dan Eccles) wrote:

> Kits for minipreps?  Sheesh.
...I don't like them either, but mainly because they are expensive and I
would like to prepare them myself...

> Anyway, helpful time!  
... and 'time is money' or more output or more papers or better job....

> We routinely use a scaled-down version of the alkaline lysis method for 
> minipreps.  If we want ultraclean minipreps (eg for sequencing) we just add a 
> PEG ppt step.  Check out my thesis for more details 
> http://www.molbiol.ox.ac.uk/www/users/rpgrant/thesis/2MaterialsandMethods.html
> and section 2.7.2
> or follow links from the bottom of this post.  The methods are very cheap, 
> dead easy, fairly robust, the only thing sacrificed is time.  However, it is 
> possible to process 40 mps at a time.

I have been using alkaline lysis + phenol for many years without major
problems, but there are a few points about it, why I would like to find
out, how to make the 'kits' with the solid resin suitable for alkaline
lysis prepared DNA myself.

1. These methods are really faster and yielded always and not only in most
cases DNA suitable for practically almost everything.
2. Phenol IS a health and environmental risk, even more chloroform,
although both probably break down to less dangerous products.

So I am still looking for some hints which resin can be used for
DNA-preparation from alkaline lysis supernatants.

Thanx
               Michael

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