Yeast DNA extraction
From 2007.igem.org
(Difference between revisions)
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-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
