Yeast DNA extraction

From 2007.igem.org

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-to leave 10 minutes 50°C
-to leave 10 minutes 50°C
   
   
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-to dissolve in 200µl TE
-to dissolve in 200µl TE

Revision as of 16:11, 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