Yeast DNA extraction

From 2007.igem.org

(Difference between revisions)
 
(2 intermediate revisions not shown)
Line 1: Line 1:
-
YEAST DNA EXTRATION
+
YEAST DNA EXTRACTION
-
-to centrifuge 5ml of inoculo 3 minutes 3000 rpm
+
-centrifuge 5ml of inoculo 3 minutes 3000 rpm
-
-to add 1ml distiled water  
+
-add 1ml distiled water  
-
-to vortex
+
-vortex
-
-to spin 20 minutes full speed
+
-spin 20 minutes full speed
-
-to risospend in 200µl SCE/zymolase
+
-risospend in 200µl SCE/zymolase
-
-to vortex
+
-vortex
-
-to leave 1 hour 37°C
+
-leave 1 hour 37°C
-
-to add 200 µl SDS
+
-add 200 µl SDS
-
-to leave 5 minutes 65°C
+
-leave 5 minutes 65°C
-
-to add 200µl 5M-Koac and to mix
+
-add 200µl 5M-Koac and to mix
-
-to leave 30 minutes in ice
+
-leave 30 minutes in ice
-
-to spin 5 minutes 14000 rpm 4°C
+
-spin 5 minutes 14000 rpm 4°C
-
-to add 200µl 5M NH4OAc and 1ml isopropanol at 500 µl surnatante
+
-add 200µl 5M NH4OAc and 1ml isopropanol at 500 µl surnatante
-
-to spin 30 minutes 3000 rpm
+
-spin 30 minutes 3000 rpm
-
-to dissolve with 90µl TE
+
-dissolve with 90µl TE
-
-to add 10µl  NH4OAc and 200µl isopropanol
+
-add 10µl  NH4OAc and 200µl isopropanol
-
-to centrifuge 1 minute 3000 rpm
+
-centrifuge 1 minute 3000 rpm
-
-to add 250µl  EtOH 80%
+
-add 250µl  EtOH 80%
-
-to spin 1 minute 3000 rpm
+
-spin 1 minute 3000 rpm
-
-to leave 10 minutes 50°C
+
-leave 10 minutes 50°C
   
   
-
 
+
-dissolve in 200µl TE
-
-to dissolve in 200µl TE
+

Latest revision as of 16:33, 25 October 2007

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