Figure 6. Semi-quantitative RT-PCR to verify the differential expression of 13 genes selected from microarrays

Each gene-specific PCR product was amplified, together with QuantumRNA 18S internal standard in the same (multiplex) PCR reaction. (QuantumRNA 18S, catalog number 1716, amplifies 488 bp 18S RNA, whereas catalog number 1717 amplifies 342 bp 18S RNA). To modulate amplification efficiency of 18S rRNA, 18S primers were mixed with 18S competimers in a 3:7 ratio. PCRs shown are representative of at least two repetitions of each PCR. Absence of mimecan, increased levels of recoverin, opsin 1, and Gng3, and decreased levels of chondroadherin, biglycan, troponin C, troponin I, hemoglobin alpha, ATPase, calcium transporting, keratin complex 1, γA-crystallin, and apolipoprotein D are observed. The expression of γC-crystallins and γD-crystallin remained unchanged. The results are consistent with those from microarray analysis.