lso tested whether sample storage affects microarray expression profiling. Duplicate mouse liver samples were dissected and either frozen at -80C for one week or stored in RNAlater at room temperature for 24 hours, 4C for one week, or -20C for one week. RNA was isolated (RiboPure, Ambion), linearly amplified (MessageAmp II aRNA Kit, Ambion), and labeled with biotin-UTP before hybridization to an Affymetrix MG 430A 2.0 array. The mean standard deviation for percent present calls for all samples was 59.4 1.1, and the correlation between samples was excellent (range, R=0.988 to R=0.996). As a representative example, Figure 1 is a scatter plot summarizing the overall results when expression levels from samples stored in RNAlater for 24 hours were compared to that of samples frozen at -80C.

Figure 1. Scatter Plot of Gene Expression Levels from Samples Stored in RNA later vs Samples Frozen at -80C. Duplicate mouse liver samples were dissected and either frozen at -80C for one week or stored in RNAlater at room temperature for 24 hours, 4C for one week, or -20C for one week. RNA was isolated (RiboPure, Ambion), and 1 g total RNA was linearly amplified (MessageAmp II aRNA Kit, Ambion). After a four hour in vitro transcription to amplify and label samples with biotin-16-UTP, the samples were hybridized to Affymetrix MG 430A 2.0 arrays (22,690 genes). This scatter plot summarizes the overall results when expression levels from samples stored in RNAlater for 24 hours were compared to that of
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