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LauraHoopes (talk | contribs) (Petite versus malonate-inhibited yeast gene expression) |
LauraHoopes (talk | contribs) (Gene Expression in Petite and Malonate-inhibited Yeast) |
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[['''GENETIC REGULATION IN EUKARYOTES, POMONA COLLEGE, 2005 PROJECT: Petite and malonate-inhibited yeast gene expression.''']] | [['''GENETIC REGULATION IN EUKARYOTES, POMONA COLLEGE, 2005 PROJECT: Petite and malonate-inhibited yeast gene expression.''']] | ||
| − | This project is an investigation of the gene expression patterns in wild type yeast, wild type yeast inhibited with malonate (an inhibitor of TCA cycle Succinate Dehydrogenase enzymes) and yeast selected to be petite. Petites have lost some or all of the mitochondrial genome and have major changes in gene expression under some culture conditions(Epstein et al, Mol Biol of the Cell 12:297-305, 2001). The class experiment explored gene expression in these three conditions using glucose as the carbon source, unlike Epstein et al., and we also hoped to study petites treated with malonate, for which little data were obtained due to a failure of amplification of RNA for the microarrays. For each condition, the class performed RNA isolation and quality control, used an Ambion kit to amplify the mRNA and couple the amplified RNA to Cy3 or Cy5, hybridized duplicate but dye-flipped GCAT arrays printed at WU, and analyzed the data using GenePix for quality control and GeneSpring for clustering and ANOVA. | + | This project is an investigation of the gene expression patterns in wild type yeast, wild type yeast inhibited with malonate (an inhibitor of TCA cycle Succinate Dehydrogenase enzymes) and yeast selected to be petite. Petites have lost some or all of the mitochondrial genome and have major changes in gene expression under some culture conditions(Epstein et al, Mol Biol of the Cell 12:297-305, 2001). The class experiment explored gene expression in these three conditions using glucose as the carbon source, unlike Epstein et al., and we also hoped to study petites treated with malonate, for which little data were obtained due to a failure of amplification of RNA for the microarrays. For each condition, the class performed RNA isolation and quality control, used an Ambion kit to amplify the mRNA and couple the amplified RNA to Cy3 or Cy5, hybridized duplicate but dye-flipped GCAT arrays printed at WU, and analyzed the data using GenePix for quality control and GeneSpring for clustering and ANOVA. We also ran RT PCR of two genes the class picked from the array data to see if we could verify our results. Results and further description can be found on the class web page by choosing the Petite Malconate Project. [http://pages.pomona.edu/~llh04747/genreg.html] |
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Laura Hoopes | Laura Hoopes | ||
Latest revision as of 00:09, 17 December 2005
'''GENETIC REGULATION IN EUKARYOTES, POMONA COLLEGE, 2005 PROJECT: Petite and malonate-inhibited yeast gene expression.'''
This project is an investigation of the gene expression patterns in wild type yeast, wild type yeast inhibited with malonate (an inhibitor of TCA cycle Succinate Dehydrogenase enzymes) and yeast selected to be petite. Petites have lost some or all of the mitochondrial genome and have major changes in gene expression under some culture conditions(Epstein et al, Mol Biol of the Cell 12:297-305, 2001). The class experiment explored gene expression in these three conditions using glucose as the carbon source, unlike Epstein et al., and we also hoped to study petites treated with malonate, for which little data were obtained due to a failure of amplification of RNA for the microarrays. For each condition, the class performed RNA isolation and quality control, used an Ambion kit to amplify the mRNA and couple the amplified RNA to Cy3 or Cy5, hybridized duplicate but dye-flipped GCAT arrays printed at WU, and analyzed the data using GenePix for quality control and GeneSpring for clustering and ANOVA. We also ran RT PCR of two genes the class picked from the array data to see if we could verify our results. Results and further description can be found on the class web page by choosing the Petite Malconate Project. [1] Laura Hoopes
