Yeast DNA extraction
From 2007.igem.org
(Difference between revisions)
Revision as of 16:10, 24 September 2007
YEAST DNA EXTRATION
-to centrifuge 5ml of inoculo 3 minutes 3000 rpm -to add 1ml distiled water -to vortex -to spin 20 minutes full speed -to risospend in 200µl SCE/zymolase -to vortex -to leave 1 hour 37°C -to add 200 µl SDS -to leave 5 minutes 65°C -to add 200µl 5M-Koac and to mix -to leave 30 minutes in ice -to spin 5 minutes 14000 rpm 4°C -to add 200µl 5M NH4OAc and 1ml isopropanol at 500 µl surnatante -to spin 30 minutes 3000 rpm -to dissolve with 90µl TE -to add 10µl NH4OAc and 200µl isopropanol -to centrifuge 1 minute 3000 rpm -to add 250µl EtOH 80% -to spin 1 minute 3000 rpm -to leave 10 minutes 50°C -to dissolve in 200µl TE
