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
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