Assessment of repeated microarray experiments using mixed tissue RNA reference samples.
Genome-scale gene expression technologies are increasingly being applied for biological research as a whole and toxicological screening in particular. In order to monitor data quality and process drift, we adopted the use of two rat-tissue mixtures (brain, liver, kidney, and testis) previously introduced as RNA reference samples. These samples were processed every time a microarray experiment was hybridized, thereby verifying the comparability of the resulting expression data for cross-study comparison. This study presents the analysis of 21 technical replicates of these two mixed-tissue samples using Affymetrix RAE230_2 GeneChip over a period of 12 months. The results show that detection sensitivity, measured by the number of present and absent sequences, is robust, and data correlation, indicated by scatter plots, varies little over time. Receiver operating characteristic (ROC) curves show the sensitivity and specificity of the current measurements are consistent with arrays previously classified as well performing. Overall, this paper shows that the inclusion of standard samples during microarray labeling and hybridization experiments is useful to benchmark the performance of microarray experiments over time and allows discovery of any process drift that, if it occurs, may confound the comparison of these datasets.
Mongan
M Ann
MA
Department of Comparative Biology and Safety Sciences, Amgen, Inc., South San Francisco.
Mongan
Ann
A
Higgins
Marnie
M
Pine
P Scott
PS
Pine
Scott
S
Afshari
Cynthia
C
Hamadeh
Hisham
H
D000818:Animals; D019540:Area Under Curve; D019985:Benchmarking / Q000379:methods; D001923:Brain Chemistry; D005069:Evaluation Studies as Topic; D020869:Gene Expression Profiling / Q000379:methods*; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009928:Organ Specificity; D012313:RNA / Q000737:chemistry* / Q000302:isolation & purification; D012372:ROC Curve; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013737:Testis / Q000737:chemistry
D012313:RNA