Yeast DNA extraction

Revision as of 16:33, 25 October 2007 by Giov1984 (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

YEAST DNA EXTRACTION

-centrifuge 5ml of inoculo 3 minutes 3000 rpm

-add 1ml distiled water

-vortex

-spin 20 minutes full speed

-risospend in 200µl SCE/zymolase

-vortex

-leave 1 hour 37°C

-add 200 µl SDS

-leave 5 minutes 65°C

-add 200µl 5M-Koac and to mix

-leave 30 minutes in ice

-spin 5 minutes 14000 rpm 4°C

-add 200µl 5M NH4OAc and 1ml isopropanol at 500 µl surnatante

-spin 30 minutes 3000 rpm

-dissolve with 90µl TE

-add 10µl NH4OAc and 200µl isopropanol

-centrifuge 1 minute 3000 rpm

-add 250µl EtOH 80%

-spin 1 minute 3000 rpm

-leave 10 minutes 50°C

-dissolve in 200µl TE