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In Vitro NER Assay Yeast extraction Buffer (200 ml): 0.2 M Tris-acetate (pH 7.5) (40 ml), 0.39 M
 

Summary: Auble Lab
In Vitro NER Assay
Reagents:
Water
YPD
Yeast extraction Buffer (200 ml): 0.2 M Tris-acetate (pH 7.5) (40 ml), 0.39 M
(NH4)2S04 (78 ml), 10 mM MgSO4 (2 ml), 20% Glycerol (40 ml), 1mM
EDTA (pH8.0) (0.4 ml), and water (39.6 ml). Store at 4C. Add DTT to
0.1 mM and protease inhibitors before use.
Protease Inhibitor Stocks: 100X PMSF (100 mM in ethanol); 1000X
Benzamidine hydrochloride (Ben) (1M in water); 1000X Pepstatin A (Pep)
(3.5 mg/ml in DMSO); 1000X Leupeptin (Leu) (1mg/ml in water); 200X
Aprotinin (Apo) (2 mg/ml in water). Store all stocks at -20C.
Solid Ammonium Sulfate
Dialysis Buffer (500 ml): 20 mM HEPES-KOH (pH 7.5) (10 ml), 20% Glycerol
(100 ml), 10 mM MgSO4 (5 ml), 10 mM EGTA (10 ml), and water (367.5
ml). Store at 4C. Add DTT to 5mM and 100X PMSF before use.
Plasmid DNA: pGEM-3Z (2.8 kbp) and pGEM-3Zf(-) (3.2 kbp). Purified by
Qiagen DNA purification kit.
N-acetoxy-2-acetylaminofluorene (NAAAF) 1 mM in ethanol. Store at -20C.

  

Source: Auble, David - Department of Biochemistry and Molecular Genetics, University of Virginia

 

Collections: Biology and Medicine