Carbos. in DNA prep!!!!!!

billing at vetsci.microvet.arizona.edu billing at vetsci.microvet.arizona.edu
Wed Aug 28 19:23:24 EST 1996


A few years ago there was a method posted here for making plasmid DNA 
from non-rough strains of E. coli.  The method was referred to as the 
"Coli Blaster".  Prior to actually making the DNA, the cell pellet was 
washed in 1.0M NaCl.  The theory behind this is that the high 
concentration of salt possibly destabilized the ion (eg., Mg)- LPS 
interactions, resulting in less LPS in the final prep.  Well, that's at 
least how it was explained to me.

It might be worth a try, assuming the CHOs are LPS, although it might 
also work for capsular material, etc.

Steve


>Joel Bader wrote:
> 
> Rodney Earl Pettway <REPE at gnv.ifas.ufl.edu> wrote:
> 
> >Could someone please help me.  I am using a DNA extraction method that
> >uses CTAB/Sorbitol/sarkosyle as detergents.  In the tube that has my
> >DNA in it there is always this bellet at the bottom.  It looks loke a
> >carbo pellet.  I has tried everything to eliminate this from my prep.
> >Please help.
> 
> >Thank you
> 
> >Rodney Earl Pettway
> I've had a similar problem and I need help figuring it out too.  Tell
> me what you find out. Thanks!
>                 Joel Bader <jabader at mindspring.com>



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