Appendix 5: Master Supply List

General Lab Equipment

• Rocker for gels (on side counter)
• Computers (1 per group)-bring a laptop
• UV/Vis spectrophotometers (in adjoining room)
• Vernier Lab Quest 2 and assorted probes (pH sensor, gas pressure sensor, dissolved oxygen sensor)
• 100 °C Heat block (in fume hood)
• Vortexer
• Carboy containing Millipore water
• Biohazard container for tip disposal-BSL1 waste
• pH meter, standards, 5 N HCl, 8 N NaOH
• Centrifuges (one large and one small)
• Balance
• Spatulas

Minimal-Use Equipment

• Microfluidizer –Mehl Core Lab. Ask TA for assistance.
• High speed centrifuge tubes (40 mL; reusable) (referred to as Oak Ridge tubes)
• SDS-PAGE Gel rigs, glass plates, and combs
• Plastic storage containers (for gels)
• 250 mL baffled flasks with purple membrane caps
• 1 L, 500 mL bottles, 150 mL bottles and flasks
• Erlenmeyer flasks
• Sterile glass culture tubes
• Shaker
• Shaker Clamps

Group

• Sterile water
• Pipette tips: 100-1000 mL, 1-200 mL,
• Falcon tube rack
• 2 group boxes (one for 4 °C and one for -20 °C freezer)
• Microcentrifuge tubes (1.7 mL and 0.65 mL) in jars
• Microcentrifuge tube rack
• Autopipette and charger
• Label tape and marker
• Ice bucket (under table)
• Scissors
• Squirt bottle of Millipore water
• Squirt bottle of 95% ethanol
• Tip disposal vessel
• Liquid disposal vessel
• Set of three micropipettes (20, 200, 1000 µL)
• Pipette stand
• Bunsen burner and tubing
• Striker

Common Drawers, Shelves, and Cabinets or Supply Bench

• 50 ml conical tubes with lids (sterile and nonsterile)
• 15 ml conical tubes with lids (sterile and nonsterile)
• 25 mL serological pipettes
• 10 mL serological pipettes
• 5 mL serological pipettes
• Styrofoam holders
• Razor blades
• Sterile Eppendorf tubes in glass jars
• Paper towels
• Gloves
• Lab Tape

Chemicals (Specialty; suppliers may be substituted)

• Autoinduction media components (adapted from Nature Protocols)
• 2X YT Broth (Fisher, BP2467-2)
• Sodium dodecyl sulfate (Fisher, BP166-100)
• Bromophenol blue, sodium salt (Sigma, B-5525)
• b-mercaptoethanol (Sigma, M7154)
• Coomassie brilliant blue G stain (Sigma, B0770)
• Coomassie (Bradford) Protein Assay reagent (Bio-Rad Quick Start)
• Lysozyme (Fisher, BP535-10)
• pH meter buffers (Thomas Scientific, numbers 4120G52, 4120G60, and 4120G68 for pH 4, 7, and 10, respectively)
• Non-canonical amino acids

Prepared Chemicals

• 30% acrylamide solution for SDS-PAGE (Bio-Rad; prepared)
• 10X running buffer for SDS-PAGE, pH 8.3 (make 500 mL)
  • o 250 mM Tris Base
  • o 92 M glycine
  • o 1% SDS
• 2X SDS load dye for SDS-PAGE, pH 6.8 (make 10 mL)
  • o 125 mM Tris HCl
  • o 4% SDS
  • o 20% glycerol
  • o 02% bromophenol blue
  • o 5% b-mercaptoethanol
• Staining solution for protein gels (TAs will prepare)
  • o 250 mL Methanol
  • o 200 mL H2O
  • o 50 mL glacial acetic acid
  • o 25 g Coomassie brilliant blue G stain
• Destaining solution for protein gels (TAs will prepare)
  • o 250 mL Methanol
  • o 200 mL H2O
  • o 50 mL glacial acetic acid
• 5 M NaCl stock
• 1 M each sodium phosphate monobasic and sodium phosphate dibasic stocks or prepare phosphate stock solution at desired pH
• 5X Equilibration for protein purification, pH 7.0
  • o 250 mM sodium phosphate pH 7.0 (refer to chart for mixing of sodium phosphate stocks to achieve 0.1 M pH 7.0 or use stock prepared at desired pH)
  • o 5 M NaCl
• 1X Equilibration/wash buffer for protein purification, pH 7.0
  • o 50 mM sodium phosphate
  • o 300 mM NaCl
• 1X elution buffer for protein purification, pH 7.0
  • o 50 mM sodium phosphate
  • o 300 mM NaCl
  • o 250 mM imidazole