BISC220/S12: Mod 2 Media Recipes
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Background Information
Lab 5: Phenotypes of Secretory Mutants
Lab 6: Analyzing Secretion Defects by Western Blotting
Lab 7: Probing and Detecting the Western Blot
Media Recipes
Yeast Series Media Recipes
| Solution | Composition |
|---|---|
| PBS (Phosphate Buffered Saline) 1X | 2.68 mM KCl 1.46 mM KH2PO4 136.9 NaCl 8.1 mM Na2HPO4 |
| PBST (PBS with Tween 20) | 1X PBS with 0.25% Tween 20 |
| YPD (Yeast Peptone Dextrose | 1% Yeast Extract 2% Peptone 2% Glucose 2% Agar (only in solid media) |
| SD (Sabouraud's Dextrose) -Ura | 0.67% Yeast Nitrogen Base 2% Glucose 0.08% -Ura amino acid drop out mix 2% Agar (only in solid media) |
| SD-Ura-His | 0.67% Yeast Nitrogen Base 2% Glucose 0.078% -Ura -His amino acid drop out mix 0.003M Histidinol 2% Agar (only in solid media) |
| Yeast 5X Protein Sample Buffer | 10% SDS 25% Glycerol 250mM Tris (pH = 6.8) 500mM DTT 0.5% BPB |
| Acrylamide Gel Tank Buffer (Gel Running) | 0.92 M Glycine 0.05M Trisma base 0.1% SDS |
| Western Blotting Buffer (Nitrocellulose Transfer) | 25mM Trisma Base 192mM Glycine 20% v/v Methanol |
| Amersham ECL Western Blotting Analysis System | www.gelifesciences.com catalog #RPN2109 |
| Strain Source | Elizabeth Valen at Swarthmore is the original source of the 3 plasmids and 3 strains Jennifer Hood-DeGrenier is the source of the Mat a strain |
