Yeast DNA extraction

(Difference between revisions)

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

@@ Line 1: / Line 1: @@
 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
--to dissolve in 200µl TE
+-dissolve in 200µl TE