Do Cells Grown in the Same Environment at the Same Time Express the Same Genes: Got cDNA?

Do Cells Grown in the Same Environment at the Same Time Express the Same Genes: Got cDNA?

Miniaci Performing Arts Center

An innovative and powerful tool for analyzing gene expression is the microarray. In this experiment, Saccharomyces cerevisiae (yeast cells) were examined to determine whether gene expression would be similar for two samples grown in the same environment. RNA was extracted and its quality and quantity was assessed. cDNA was made from the mRNA (taken from the total RNA sample) and its subsequent presence was verified by amplifying a known gene that should always be expressed, TDH1, and visualizing it on an agarose gel. The cDNA samples were labeled with the appropriate fluorescent dyes and hybridized to the array. The slide was rinsed, dried, and scanned by a computer and found to contain uniform dye incorporation on each spot. The results indicate that indeed, cells grown in the same environment at the same time do express the same genes. Valuable information was obtained from this experiment regarding reliable protocols that can be used to consistently isolate good quality total RNA, make labeled cDNA from the mRNA, and hybridize the cDNA on to a microarray slide for analysis. Future applications include using the microarray tool to determine the difference in gene expression with yeast exposed to various conditions and during different life stages of the cell.