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TEN-MINUTE GENOMIC DNA PREPARATION -Sterile water
 

Summary: Auble Lab
TEN-MINUTE GENOMIC DNA PREPARATION
FROM YEAST
Reagents:
-YPD
-Sterile water
-Lysis buffer: 2% Triton X-100, 1% SDS, 100 mM NaCl, 10mM Tris-HCl (pH 8.0),
1mM Na2EDTA.
-Phenol:Chloroform:isoamyl alcohol (25:24:1).
-Glass beads: 0.45-0.5 mm beads (Sigma) soaked in nitric acid and washed in
distilled water. Beads should be dried before use.
-TE (pH 8.0): 10 mM Tris-HCl, 1 mM Na2EDTA
-100% EtOH
-Rnase A (10 mg/ml)
-7.5 M ammonium acetate
Protocol:
1. Grow 10-ml yeast culture to saturation in YPD at 30C.
2. Collect the cells by centrifugation for two minutes in a clinical centrifuge.
Remove the supernatant and resuspend the cells in 0.5 ml of distilled
water. Transfer the cells to a 1.5-ml microfuge tube and collect them by

  

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

 

Collections: Biology and Medicine