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Since we did not suspect any problems with the randomization, no baseline tests of imbalance were performed [ 32].
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The normalized expression of the two replicates were combined and tested for significance of imbalance using binomial and chi square tests.
When applied to allele specific counts on a gene, rather than SNP by SNP level, this approach effectively tests allelic imbalance of gene expression.
AA-ALIGNER was designed to be modular allowing for allowing for the incorporation of alternative methods for variant identification and tests for significance of imbalances.
For most of our results, we required a minimum of five reads per allele when testing for imbalances to prevent erroneous testing of homozygous variants.
Unlike the results from our cross-validation tests, sample imbalance had more of an effect in these independent data set tests.
P χ2: Chi-square test (Pearson P-value); 95%CI: 95% Confidence Interval * degreegree of imbalance) in module; ** R2 (degree of correlation).
A crisis of imbalance.
The clinical significance of remaining imbalances was assessed by examining the functional content of the region of imbalance, referencing known benign and pathogenic CNVs (ISCA, DECPIHER, OMIM) and an internal clinical database of previously tested individuals (Moka).
Higher ERI indicates higher level of imbalance.
We experimentally detected differential protein binding at six of nine tested imbalance predictions from AA-ALIGNER for CREB1 (Cyclic-AMP Responsive Element Binding protein 1) binding in GM12878 ChIP-seq data, including imbalances at two disease-associated loci.
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