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For a detailed description of the selectivity and specificity checks see previous reports [29], [30].
Therefore, the specificity checks confirmed that the TFs involved in the prediction were related to CLL pathology.
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Primer-BLAST allows for specificity check of the selected primers against the set of reference sequences.
First-strand cDNA synthesis products were diluted fivefold, and one μl of cDNA was used to carry out semi-quantitative RT-PCR for a primer specificity check.
The default settings were kept for all other parameters and no specificity check was done between transcripts within the same cluster.
Primers were selected based on stringent criteria, in particular with regard to the sequence specificity check step (details described in Wang and Seed 2003).
Primers were designed using Primer3Plus software for real-time PCR [ 50], and specificity checked using Primer-BLAST against NCBI A. pisum RefSeq gene models (primer sequences are shown in Additional file 1: Table S1).
Species-specificity checking against 15 important meat and fish and 5 plant species detected no cross-species amplification.
Assay specificity was checked using cytoblocks prepared from cultured cells with siRNA knockdown (Supplementary Figure 2).
The assay specificity was checked using control patients with a variety of urological disorders.
These influenza-specific cell lines contained 15 to 30% tetramer HLA-A2/Flu58-66 pospecificpecific T cells; their specificity was checked using irrelevant and Flu-peptide pulsed T2 cells.
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