Smolke:Protocols

Protocols

Cloning

• Primer Design
• Plasmid prep with Epoch columns and homemade buffers
• PCR amplifying inserts from plasmids
• When to use phenol-chloroform vs. columns
• Restriction digests
• Gel extraction
• Ligation
• Transformation
• Colony PCR
• Screening by digestion
• Sequencing with Elim
• Making freezer stocks
• Gene assembly

Yeast

Protocols

• Pouring yeast dropout plates
• Yeast plasmid transformation
• Yeast colony PCR
• Anti-V5 Western
• Library-scale yeast transformations
• Yeast chromosomal modifications
• Yeast plasmid prep
• Yeast metabolite extraction

Observations and Discussion

• CSY3

Tissue Culture room

• Adherent cell care and feeding
• Freezer stock management and database
• General maintenance

Miscellaneous

• Glass Beads
• Poster Printing
• My pipet-aid is broken!
• Clearing Quanta Clogs
• Cleaning Bottles for the LC

Solutions

• Pfu Buffer
• Taq Buffer
• 10x DO Media
• dNTP Mixture
• 5-Fluoroorotic acid
• Antibiotic Stocks