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In publically available data sets, interrogated using Oncomine, RPL3 was downregulated: two-fold in ovarian adenocarcinoma compared with normal ovary (P-value 6.94 × 10−10) (Welsh et al, 2001), and 36-fold in breast tumour stroma compared with normal stroma (P-value 1.06 × 10−28) (Finak et al, 2008).
The two independent data sets interrogated were generated previously from normal human monocytes purified from fresh blood samples from 1370 individuals [Gutenberg Health Study (GHS) (16)] and 753 individuals [Cardiogenics Project (CGP) (17), http://www.cardiogenics.eu/web/], respectively, and subjected to genome-wide genotyping and microarray gene expression analysis.
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Similarly, it is not clear if these "mega probe sets" have excluded probes from problematic probe sets, such as the probe sets interrogating incorrect strand as described for Surf4 [3].
The consistency of the P/A calls between probe sets interrogating the same gene provides an indicator of how differently the redundant probe sets from the same gene behave at this level.
When we inspected the gene structures and probe set consensus sequence alignments using the Integrated Genome Browser, we found that about half of our findings have redundant probe sets interrogating different target mRNAs based on the knownGene transcript forms.
For the purposes of redundant probe set assignments and examining alternative splicing/polyadenylation, it is better to examine the exon-intron structure of target mRNAs via their genomic alignments, since ultimately these are what biologists and data analysts will use to assess whether a given pair of probes sets interrogate an alternatively spliced or polyadenylated gene.
This cut-off, although relatively high, was optimized to reduce the occurrence of false negative data in data sets interrogating a small number of gene sets.
The U133 Plus 2.0 array provides over 54,000 probe sets interrogating the human genome and over 39,000 of the probe sets measure CNVs directly within transcribed genes (Table 1).
The refFlat.txt.gz file[ 29] contains physical positions of gene locations according to the human genome assembly version Build 35 and has been used for identification of probe sets interrogating genes.
Apart from this, methodological issues such as the different primer sets interrogating methylation at distinct CpG dinucleotides of a specific promoter region could explain the range of frequencies reported in the literature.
In the second experiment, a commercially available Affymetrix Drosophila full genome array was used (Drosophila Genome Array, Affymetrix) which contains probe sets interrogating more than 13,500 genes from Drosophila melanogaster (experiment B).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com