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The normalization and data filtering resulted in 1088 high quality probe signals, which represented both human and porcine miRNAs.
For each 2 bp position, s, in a unigene covered by at least one probe, we calculated the single position polymorphism deviation (SPPdev) as: SFPde v s = Σ w k SFPde v k, Where w k is the weighting factor for the high quality probe k that covers position s in the unigene.
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When choosing a small minimum inter-probe distance (≪probe-length) for the intergenic regions a "selective tiling" is achieved, i.e. high density, but with high quality probes only (see Table S1 for maximum probe density).
From the 411 probes with normal distribution in both sets as well as in the combined set, 131 failed the range criterion leaving 280 high quality probes in 250 genes for further study of heritability, age and gender effects.
The high quality probes were defined as those probes whose signal intensities exceeded those of a defined percentile of anti-genomic probes.
The main limitation is the design of high quality probes sets to expect a full coverage of 16S rDNA sequences as complete as possible.
The initial set of 15,875 high quality probes with a cross hybridization potential of zero were used to query SWISPROT with BLASTx.
To design high quality probes, we developed the CADO4MI program which allows a validation of oligonucleotides by cross-comparison of their sequences with data from several reference databases.
Deviation in hybridization intensities was calculated using a 2 bp sliding window incorporating data from all high quality probes covering each position.
High quality probes with coronal data (165 probes with structures in 5' UTRs, 1,188 probes in 3' UTRs and 66 probes in both UTRs; see Methods for selection criteria) are clustered in modules of correlated expression [ 52].
We similarly restricted our DNA methylation analyses to a set of 335,307 high quality probes with a high degree of sequence conservation between humans and chimpanzees (as in Hernando-Herraez et al., 2013; see 'Materials and methods').
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