Table 3.

Real Time PCR results confirming differential expression identified by microarray analysis

Clone ID	Forward Primer	Reverse Primer	Federated Ratio	Fold Change Identified by Real Time RT PCR	Fe Efficient Standard Error	Fe Inefficient Standard Error
Gm-c1028-4867	CAGTGGAACTTCGTTGGG	AAAAGGCCTGGAATGCTC	2.379	7.56	0.345	0.255
Gm-c1004-8188	CCCTGATCTAGAAGTTGG	GCAGGAGCAGATGGTAGC	2.412	2.9	0.185	0.015
Gm-c1028-5360	CAGTGGAACTTCGTTGGG	AAAAGGCCTGGAATGCTC	2.701	2.7	0.115	0.030
Gm-c1004-5020	GAAGAACAGCGAAACCTAAC	CGGCTACTCCCTATCCA	2.772	2.7	0.020	0.040
Gm-c1028-2326	CAAGAGCATGATCTACCAGC	GGACAGAGGGAGAGATCAGG	3.156	2.82	0.080	0.040
Gm-c1013-2943	CGAACCCAAACAAGATACAC	GATTGTATTTCCCGTGGATT	3.453	5.12	0.040	0.060
Gm-c1028-8658	TCCAACTCCATCGTCGAG	GTGAATGCGCGAAGGAT	3.564	4.2	0.055	0.010
Gm-c1028-8183	CCAAGCTGGACCATA	ACATTGGCTATTTACTTACA	3.687	3.66	0.025	0.045
Gm-c1028-963	TGCCATCACTGTTTATCAAG	GCCACTGCCCTGTCTTACTC	7.149	2.8	0.060	0.05

Ten genes were chosen at random to have their differential expression between Clark (Fe-efficient) and IsoClark (Fe-inefficient) identified by microarray analysis confirmed by semi-quantitative real time PCR analysis. Differential expression was confirmed for nine of the ten genes chosen. In four of the nine genes the real time PCR showed greater differential expression between the NILs than was identified by microarray analysis.