Yeast DNA extraction

From 2007.igem.org

(Difference between revisions)
Line 2: Line 2:
-to centrifuge 5ml of inoculo 3 minutes 3000 rpm
-to centrifuge 5ml of inoculo 3 minutes 3000 rpm
 +
-to add 1ml distiled water  
-to add 1ml distiled water  
 +
-to vortex
-to vortex
 +
-to spin 20 minutes full speed
-to spin 20 minutes full speed
 +
-to risospend in 200µl SCE/zymolase
-to risospend in 200µl SCE/zymolase
 +
-to vortex
-to vortex
 +
-to leave 1 hour 37°C
-to leave 1 hour 37°C
 +
-to add 200 µl SDS
-to add 200 µl SDS
 +
-to leave 5 minutes 65°C
-to leave 5 minutes 65°C
 +
-to add 200µl 5M-Koac and to mix
-to add 200µl 5M-Koac and to mix
 +
-to leave 30 minutes in ice
-to leave 30 minutes in ice
 +
-to spin 5 minutes 14000 rpm 4°C
-to spin 5 minutes 14000 rpm 4°C
 +
-to add 200µl 5M NH4OAc and 1ml isopropanol at 500 µl surnatante
-to add 200µl 5M NH4OAc and 1ml isopropanol at 500 µl surnatante
 +
-to spin 30 minutes 3000 rpm
-to spin 30 minutes 3000 rpm
 +
-to dissolve with 90µl TE
-to dissolve with 90µl TE
 +
-to add 10µl  NH4OAc and 200µl isopropanol
-to add 10µl  NH4OAc and 200µl isopropanol
 +
-to centrifuge 1 minute 3000 rpm
-to centrifuge 1 minute 3000 rpm
 +
-to add 250µl  EtOH 80%
-to add 250µl  EtOH 80%
 +
-to spin 1 minute 3000 rpm
-to spin 1 minute 3000 rpm
-
-to leave 10 minutes 50°C  
+
 
 +
-to leave 10 minutes 50°C
 +
 +
 
-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