Plasmid DNA miniprep

[Manuel G. CLAROS]

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__________________________________
|   Ecole Normale Superieure - CNRS    |    E-mail: claros at biologie.ens.fr    |
|   Lab. Genetique Moleculaire         |    Fax:    331-44 32 39 41           |
|   46 rue d'Ulm                       |    Phone:  331-44 32 39 39           |
|   75005 Paris (France)               |                                      |
_______________________________________________________________________________