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In order to generate a human breast tumor predictor, we utilized the list of significantly differentially expressed genes identified previously from the mouse gene expression data.
We transformed and normalized mouse gene expression data as described4.
RNA amplified by Nugen kit was hybridized on Affymetrix MOE 430 2.0 mouse gene expression microarrays.
Zhang, W. et al. The functional landscape of mouse gene expression.
T.I., M.T. and A.M. contributed to the design of the mouse gene expression experiments.
Pritchard, C.C., Hsu, L., Delrow, J. & Nelson, P.S. Project normal: Defining normal variance in mouse gene expression.
Microarray analysis was performed by TakaraBio using SurePrint G3 Mouse Gene Expression 8 × 60K (Agilent, Santa Clara, CA, USA).
Purified cDNA was fragmented to uniform size and applied to Agilent Mouse Gene Expression 4 × 44 v2 Microarray (Agilent Technologies, design ID 026655) in hybridization buffer.
Analysis of mouse gene expression levels was carried out using the Sybr green I master mix (Roche), with the following primers: gapdh_F: 5′-CATGGCCTTCCGTGTTCCTA-3′, gapdh_R: 5′-GCGGCACGTCAGATCCA-3′ Dusp1_F: 5′-GTGCCTGACAGTGCAGAATC-3′, Dusp1_R: 5′-CACTGCCCAGGTACAGGAAG-3′, Il33R_F: 5′-AGACCTGTTACCTGGGCAAG-3′, Il33R_R: 5′-CACCTGTCTTCTGCTATTCTGG-3′.
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For the study presented here, we performed a comparative analysis of human-mouse gene expression patterns to assess the extent of expression divergence between the two species and to explore the connections between the evolution of gene expression and function.
aGEM v2.0 is a powerful Platform that integrates several important mouse gene-expression resources.
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