Freimoser:Protocols:FastYeastTransformation
From OpenWetWare
(Difference between revisions)
| Line 11: | Line 11: | ||
__NOTOC__ | __NOTOC__ | ||
| - | == Protocol: | + | == Protocol: Fast yeast transformation== |
#50ul carrier DNA | #50ul carrier DNA | ||
Revision as of 08:11, 17 October 2006
Protocol: Fast yeast transformation
- 50ul carrier DNA
- scrap cells from plate
- 1-2ul DNA-plasmid (up to 30-50 ul)
- 240 ul PEG (50%)
- 36 ul Li-Ac (1M)
- resuspend with blue tip
- incubate at 30°C or RT at least 30 min
- heat shock: 45°C, 15 min (or 42°C for 20 min)
- spin down, resuspend in 100ul H2O and plate everything on corresponding medium
Material:
- Plasmid: 100 ng-1ug
- carrier DNA: salmon/herring sperm DNA in H2O, 2mg/ml stock, 500 ul aliquots, heat-inactivated at 95°C for 5-6 min and put on ice before use (has to be done once).
- 50% poly-ethylen-glycol (PEG) (MW 33-50) solution, filter-sterilized
- 1M Li-Acetate, autoclaved
