Results

Yeast Culture

Yeast culture growth was monitored by absorbance to obtain healthy yeast at optimal exponential log phase.  Table 1 shows absorbance after over night growth.  Culture was diluted with media in order to obtain equal concentrations of cells for the RNA extraction. 

Table 1

RNA isolation

RNA isolation resulted in relatively pure RNA based on the 260/280 absorbance ratio (Table 2).  The RNA was also concentrated enough to obtain 8 ug for hybridization.  ΔZMS2 was at 625.8 ug/ul and WT was at 654.4 ug.ul.  The RNA was run on a gel to determine it's integrity (Figure 1).  Lane 1, containing ΔZMS1, contained DNA as well as a band for the 28s subunit.  Lane two, containing WT, contained bands for 28s, 18s and 5s.  Bands were visible but the RNA was too degraded to yield promising results.  Tuesday Group 2 wild type  and Group 4 ΔZMS1 was sused instead.

Table 2

Figure 1.  Gel of RNA extraction.  Lane one is ΔZMS1 and lane 2 is ΔZMS2.

Microarray Slide

The microarray slide was broken when shipping.  Slide 726 from previous years was used for analysis instead. 

Figure 2.  Microarray scan showing DNA spots for yeast.

Genes of Interest

GCAT website provided information on specific genes (Campbell 2009).  Genes that were related to oxidative stress (Table 4).  Eight genes that have a p-value less than 0.01 (alpha) were chosen for further analysis and considered differentially expressed (Table 5).  Uga2 was chosen for further analysis because of its presence in the glutamate catabolic pathway, role in reducing NADP and its statistical significance.

Table 4

Table 5

Function from http://yeastgenome.org/

Link to Discussion