Protocols used by the Peccoud Lab: molecular biology, DNA synthesis, DNA sequencing, yeast genetics, and imaging protocols.

Creation of 0.5% Zinc Acetate

Materials Consumables: Zinc acetate DI water Bottle top filter Hardware: 250 ml beaker or erlenmeyer flask Stir bar Stir plate Sterile bottle Instructions  Add 0.5 g zinc acetate to 100 ml deionized water while mixing with stir bar in beaker.  Filter sterilize using bottle top filter (0.2 µm) into sterile bottle. [...]

Creation of 0.5% Zinc Acetate2019-08-22T22:02:17+00:00

Creation of 1.5 M Tris pH 8.8 Buffer

MaterialsConsumables:Tris(hydroxymethyl)aminomethane (Tris) 181.65 gDI WaterConcentrated HCIHardware:MixerStir barAnalytical balanceGraduated cylinderSterile 1L bottleInstructions Add 181.65 g of Tris to a 1L bottle with a stir bar Add 800 ml of DI water and mix thoroughly to dissolve Add HCI until pH of 8.8 is achieved Add remaining DI water so a final volume of 1L is reachedNote: Make sure to let [...]

Creation of 1.5 M Tris pH 8.8 Buffer2019-08-22T21:56:10+00:00

Isolation of Plasmids via Zymo Research Mini-Prep

Getting Started Before proceeding: inoculate liquid media and grow sample using the Culturing E. coli protocol.     Materials Consumables: Binding buffer II-P spin column Microcentrifuge tubes Ice Zymo Research Mini-Prep Kit Hardware: Centrifuge Procedure  Set up all the tubes necessary for the mini-prep in a tube rack and number all the tubes (i.e. [...]

Isolation of Plasmids via Zymo Research Mini-Prep2019-08-22T21:19:46+00:00

DNA Fragment Analysis via 4200 Agilent TapeStation

Materials Consumables: D5000/Genomic Sample Buffer D5000/Genomic Ladder Strip Tubes/96 well plate Strip Caps/Aluminum Foil Seal D5000/Genomic Screentape Hardware: 4200 TapeStation Agilent Computer Vortexer Prepping Samples Launch the '4200 TapeStation Controller' Software Controller Software Interface Assign a location to each samples you will be analyzing in the digital layout of the wells Select the wells [...]

DNA Fragment Analysis via 4200 Agilent TapeStation2019-07-08T18:59:38+00:00

YPD Agar Growth Media

Materials Consumables for 1L of media: 900 mL Distilled Water 20 g Peptone 10 g Yeast Extract 20 g Agar 100 mL (bottle) 20% glucose Plastic Weigh Boat Aluminum Foil Autoclave Tape 0.2μm filter 3 mL syringe Hardware: 2L Flask Stir bar Hot plate Stir bar magnet stick Autoclave tub Autoclave Media [...]

YPD Agar Growth Media2019-08-22T21:09:16+00:00

TE Buffer

Materials Consumables: Tris(hydroxymethyl)aminomethane (Tris) 0.6057 g Disodium Ethylenedeamine tetraacetate.2H2O (EDTA) 0.1861 g HCI Hardware: Mixer Stir bar Analytical balance Graduated cylinder 500 ML bottle Autoclave Prepare Buffer  Add 0.6057 g of Tris and 0.1861 g of EDTA to a 500 mL bottle with a stir bar  Add 450 ml of DI water and mix thoroughly [...]

TE Buffer2019-07-22T21:25:07+00:00

Chloramphenicol Antibiotic Stock

Materials Consumables: Chloramphenicol (323.13 g mol -1) Ethanol (96%) 50mL conical vials Screw cap tubes Hardware: Vortexer Analytical balance Procedure Instructions Pipet 40mL of ethanol into a 50mL conical vial Weigh out 2g of chloramphenicol with the analytical balance Pour into ethanol and vortex Once fully dissolved, filter sterilize: Screw 0.2μm filter onto 3mL [...]

Chloramphenicol Antibiotic Stock2019-07-22T21:18:44+00:00

Carbenicillin Antibiotic Stock

Materials Consumables: Carbenicillin (Sodium salt; 371.39 g mol -1) Screw cap tubes 25mL Seriological Pipet 50mL Conical vial in a tube rack DI Water Hardware: Vortexer Electric serological pipet Dissolving Antibiotic Creation of 500mg/uL Carbenicillin Stock Weigh out 10g of Carbenicillin Place in 50mL conical vial Pipette 15mL of DI water into conical vial [...]

Carbenicillin Antibiotic Stock2019-07-22T21:12:24+00:00