NH4Ac-Miniprep becomes bad - why?

H. ROYCHOWDHURY 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
> culture. 
> 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
> reoccures.
> 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???
> 
> Thanks
> Mike
> 
> 
> 
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 
a month.

			>>>>>>>>>>>>>>>>>>>>>>>>>>>
			  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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