now you see it now you don't

Cubano67.ix.netcom.com Cubano67.ix.netcom.com
Wed Nov 8 20:12:06 EST 1995


ESA009 at ED.SAC.AC.UK ("Dr Rob Harling") wrote:

>We're using RAPD-PCR to study diversity in epiphytic Pseudomonas 
>species.  We prepare the template DNA using Flowgen's Puregene kit, 
>which is basically an SDS cell lysis, ammonium acetate protein 
>precipitation, RNAse, isopropanol DNA precipitation, then dissolve in 
>water.  The resulting DNA is of good spectrophotometric quality. 

>PCR works if we use freshly - prepared DNA, ie extracted from the 
>bacteria then run on the same day.  If we store the DNA at 4C for 
>only one week, then PCR fails, we get zilch in the lanes at any 
>concentration of template.  The DNA still looks the same after one week
>when run on a gel, ie there doesn't seem to be nuclease breakdown
>of the DNA.  Does anyone know what's happening to the DNA in storage?

What is your storage buffer (if any) and why do you store at 4C?

>Thanks
>Rob (still got my hair) Harling

>Dr Rob Harling
>SAC (Scottish Agricultural College)/
>  University of Edinburgh
>West Mains Road
>Edinburgh EH9 3JG
>Scotland, UK
>tel: +44 (0)131 535 4000
>fax: +44 (0)131 667 2601
>e mail: esa009 at ed.sac.ac.uk






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