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Raw probe summary data was exported into Microsoft Excel and transformed on the log base 2 scale.
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78 (or 7.3%) of HuEx 1.0 and 11 (or 3.6%) U133Plus2 probes with sequence complementary to target did not respond to changes in concentrations; see Supporting Tables S1 and S2 for probe summaries.
Probe summaries were calculated in BeadStudio and quantile normalised.
The probe summaries were generated by using RMA from the BioConductor collection of packages.
Li-Wong model based index was used as probe set summary measure [107].
The model-based Robust Multi-array Average (RMA) algorithm was used to generate the probe set summary based on the full annotation on gene- and exon-level [31].A filtering step was introduced and only genes that are expressed above a defined intensity threshold of 132.5 on 10% of the arrays were retained, i.e. at least on 3 of 28 arrays.
Tests based on expression indices were carried out on the RMA probe set summary values.
A probe level summary was determined using the Affymetrix GeneChip Operating Software using the Microarray Analysis Software 5 algorithm.
The RMA probe set summary algorithm uses only PM values and employs RMA background correction and quantiles normalization.
We used RMA [ 49] to obtain the probe set summary values for the Affymetrix U133A and U133Plus_2 Genechips.
We acknowledge that, in all likelihood, testing methods based on probe set summary values would have exceeded the performance of the methods based on probe level tests had a probe set level normalization been performed for the Golden Spike data.
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