The end of the World

Andre Hamel hamel at ccu.umanitoba.ca
Tue May 18 09:27:15 EST 1993


>Has anyone else out there had trouble sequencing directly from DNA prepared
> with Promega's Magic Maxipreps? I have found that the preps seem OK by 
>restriction analysis, but will give no sequence by Sequenase double stranded 
>sequencing even after repeated phenol extractions. The same DNA when 'crudely'
> prepped by standard miniprep procedure gave fine sequencing results, so it
> is a fault with the magic maxi. Other people in my Lab have had similar results.
>
>Mike Poidinger
>Dept of Microbiology
>University of Reading
>Et tu Kitus?

I haven't used their Maxi prep kits, only the mini one ... I found it
necessary to elute the DNA (after washing/drying column) with HOT water
(80oC), directly into NaOH such that final conc. is 0.2 N ... then
incubate at least 37oC for 15 min ... add NaOAc, pH 4.5 to 0.3 M final
conc., then 4 times vol. etoh ... </= -20oC, etc.

cheers
   ----------------------------------------------------------------------
Andre Hamel                                 email: hamel at ccu.umanitoba.ca
Manitoba Veterinary Services                lab tel.: (204) 945-7630
Infectious & Genetic Disease                home tel.: (204) 275-1204
Mol.Biol.Lab                                FAX: (204) 945-8062
545 University Crescent                   
Winnipeg, Manitoba                          
CANADA
R3T 5S6
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