Abstract       Introduction        Materials and Methods       Results      Discussion      Sources Cited

All the procedures for the RNA isolation, probe construction, microarray hybridization and analysis can be found at James Madison University’s BIO 480/580 Lab course website.

Probe Construction and Hybridization

096 (WT green, ZMS1∆ red)

Total RNA was isolated and purified from the other cellular components from yeast strains, ZMS1∆, ZMS2∆, and wild type yeast, grown in liquid culture for use in the microarray analysis. 

From the purified RNA, cDNA probes were constructed. The mRNA was reverse transcribed with poly T-primers with a 3DNA capture sequence for a specific dye. The Wild Type (WT) yeast cDNA was labeled with green dye and the ZMS1∆ cDNA was labeled with red dye. 

            Microarray Slide 096 was subjected to a series of hybridization and wash steps to anneal the cDNA probes to the 70mer sequences printed on the slide and to anneal the dyes to the 3DNA capture sequences.

Data analysis

725    (WT red, ZMS2∆ green)

726    (WT green, ZMS1∆ red)

695    (WT green, ZMS2∆ red)

  *Microarrays used for analysis were obtained from Fall 2008.

Slide 725 was analyzed due to lack of results. On this slide genes from zms2∆ cells were labeled with green and genes from WT cells were labeled with red. To analyze the microarray data the slides were gridded and segmented using Scanalyze. The numerical data was transformed to log base two and standardized with the mean set to zero and the standard deviation set to one using Magic Tool. To find genes which are up or down regulated six housekeeping genes were averaged to calibrate what an up or down regulated gene means. Above one and below negative one were determined to be up or down regulated.