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This provided a high confidence dataset of 434 chips on which a final background correction, normalization, and cluster analysis for all the data were performed.
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Normalization and clustering were performed in GeneSpring software (Agilent Technologies).
This paper breaks new ground in normalization and clustering methods for highly replicated temporal microarray data.
Multi-array normalization and clustering analysis were performed using a hierarchical method, with an average linkage and Euclidean distance metric.
The hybridization signal was subjected to normalization and clustering by using open-source R (ver. 2.1.1) packages and S-Plus (Insightful, WA).
(A ) Normalization and clustering were performed according to the procedures described for Figure 3C legend.
(XLSX 62 kb) Additional file 12: Comprises of additional details of the methods, including the options and parameters of the tools used for processing of the reads, known miRNA expression profile generation, normalization and clustering and novel miRNA prediction pipeline.
Then data normalization by a normalization factor and clustering was performed based on mean intensity for inter-array comparison.
Quality control analysis, data normalization and hierarchical cluster were performed as described.
Though other inter-platform normalization methods and clustering methods were tested (z-score, quantile normalization), we found that only by using a common reference sample and transforming single intensity data into a log ratio enabled us to overcome the platform-dependent bias and observe cross-platform cell line-specific clusters.
Intensity-dependent print-tip MAD normalization, Quantile normalization, and hierarchical clustering [63] were performed with R as described before [64].
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