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
• Electrocompetent Transformation
• Heat Shock Transformation
• Colony PCR
• Screening by digestion
• Sequencing with Elim
• Making freezer stocks
• Gene assembly
• EtBr stock

Yeast

Protocols

• Yeast Media Recipes
• Pouring yeast dropout plates
• Yeast plasmid transformation
• Quick Yeast plasmid transformation
• Yeast colony PCR
• Anti-V5 Western
• Library-scale yeast transformations & Gap repair
• Yeast chromosomal modifications
• Yeast plasmid prep
• Feeding yeast cultures
• Yeast metabolite extraction
• Heme measurements

Observations and Discussion

• CSY3

Tissue Culture room

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

Quanta

• Preventing Clogs
• Clearing Clogs

VYB

• Guidelines

Miscellaneous

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

Solutions

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