Plasmid DNA miniprep

Manuel G. CLAROS claros at biologie.ens.fr
Wed Jan 19 09:51:43 EST 1994


In Article <2hjp7p$alt at news.u.washington.edu>, findley at u.washington.edu
(Seth Findley) wrote:
>
>I would greatly appreciate any references for the most current (and easy):
>
>
>Plasmid Miniprep Protocol for 1-10mls from Yeast (C. c.)
>
>Thanks, in advance.
>
1 ml culture on YEPD, SD (or a big colony in a plate)
Spin and resuspend cells in 100 ul of
        50 mM Tris pH 7,5
        10 mM EDTA
        0,3% bME
        1 mg/ml zymoliase T100
Incubate 1h at 37C
Add 20 ul SDS 10%. Mix up to lysis
Add 100 ul AcNH4 7M
Precipitate 15 min at -80C. Spin 15 min 15K
Take supernatant and add 0,7 vol isopropanol
Precipitate 15 min -80C. Spin 15 min 15K
Rinse pellet with 1 ml ethanol 70% RT. Spin. Dry
Resuspend pellet in 20 ul water.
Transform 100 ul E.coli with 5 ul DNA. ATTENTION, let 2 hours for expression!

I hope this will help

Cheers

Manuel

_____________________________Manuel_G._CLAROS__________________________________
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