Yeast DNA extraction
From 2007.igem.org
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