Microarray analysis for Saccharomyces cerevisiae

J Vis Exp. 2011 Apr 7:(50):2585. doi: 10.3791/2585.

Abstract

In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen peroxide (H₂O₂), an oxidizing agent. In the experiment, yeast is grown for 48 hours in 1/2X YPD broth containing 3X glucose. The culture is split into a control and treated group. The experiment culture is treated with 0.5 mM H₂O₂ in Hanks Buffered Saline (HBSS) for 1 hour. The control culture is treated with HBSS only. Total RNA is extracted from both cultures and is converted to a biotin-labeled cRNA product through a multistep process. The final synthesis product is taken back to the UVM Microarray Core Facility and hybridized to the Affymetrix yeast GeneChips. The resulting gene expression data are uploaded into bioinformatics data analysis software.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Hydrogen Peroxide / pharmacology
  • Oligonucleotide Array Sequence Analysis / methods*
  • Oxidative Stress / drug effects
  • Oxidative Stress / genetics
  • RNA, Fungal / analysis
  • RNA, Fungal / genetics*
  • RNA, Fungal / isolation & purification
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism

Substances

  • RNA, Fungal
  • Hydrogen Peroxide