NH4Ac-Miniprep becomes bad - why?
hroychow at NMSU.EDU
Thu Aug 29 10:24:13 EST 1996
On Thu, 29 Aug 1996, * wrote:
> Hi there
> Recently I switched from the Qiagen miniprep protocol to an
> alternative method using an RNase containing resuspension buffer as
> well, lysing with NaOH/SDS but precipitating genomic DNA and proteins
> with 6M Ammoniumacetate. Precipitating the supernatant with Isoprop
> gave a yield of plasmid (sequencable) as high as 25ug from a 1.5mL
> So I made a kit containing all the components that I need for the
> prep. After a while (month) though my yields decreased rapidly until
> the method didn't work at all anymore. So I made fresh solutions and
> *bingo* it woked again. Now, after another month, the same problem
> It would be much easier if I knew *what* solution is becoming bad and
> I would be interested in *why* it happens and how to *avoid* it.
> Anybody with similar problems???
Just a guess... Do you store your Amm.Acetate soution at room
Temperature? In that case, you do not have much of it left in soln. after
Dr. Hiranya S. Roychowdhury
Plant Genetic Engineering Lab.
Box 3GL, NM State Univ.
Las Cruces, NM 88003
Phone: (505) 646-5785
hroychow at nmsu.edu
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