Team:HKUST/Protocols/GFP testing

Fluorescence microscopy visualization

Procedure:

1. Grow yeast transformed cells containing appropriate plasmids in 5 ml SCM minus His & glucose at 30ºC overnight.
2. Add galactose (to a conc.of 2%) at a cell density corresponding to an OD600 of a 10-fold dilution reaching 0.8.
3. Continue growing cells at 30ºC for 30min to 1.5hr.
4. At time 30min, 60min, 90min, use 1ml of cells, spin down and resuspend cells in 10μl ddH2O.
5. Use 2μl cells and visualize the cells in a fluorescency microscopy.

• Agarose gel preparation and gel electrophoresis
• PCR (Taq, Vent)
• PCR product clean-up
• Enzyme digestion (vector, insert)
• Plasmid DNA extraction
• Gel cutting
• Gel extraction
• Ethanol precipitation
• Ligation
• Transformation to E.coli
• Transformation to yeast
• Cell lysis
• Yeast genomic DNA extraction
• Western blotting
• GFP testing