Design of Yeast Genomic Expression Experiments-试验方案

Pat Brown's Lab ,Department of Biochemistry and the Howard Hughes Medical Institute

I. Experimental setup

A. Choose the experimental parameters

1. Choosing the strain background: Many labs routinely work with their favorite

strains, but it is worthwhile to consider which strain is best for your genomic

studies, because different strains can have very different genotypes and

phenotypes

a. Be aware of the differences between common lab strains (eg. S288C vs

W303, etc.) and how those differences might the experimental results

b. The mating type can affect phenotype, including gene expression

c. Best to use strains with minimal auxotrophic markers to simplify analysis of

the results

2. Choosing the experimental conditions

a. Determine the optimal media, culture type, temperature for your needs

b. Investigate the optimal “dosage” of the stimulus: Many genomic

studies observe the response of cells to various stimuli. It is worth

investigating the dosage of the applied stimulus to pick conditions that

will give meaningful results

1) Lethal conditions may result in data that are difficult to interpret,

while mild conditions may not provoke a detectible genomic

expression program

c. Investigate the appropriate time points for each timecourse experiment