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Additional analysis of secondary and somatic analysis tools based on the simulated NA12878 genome are provided in the Supplementary Material.
Somatic analysis detected mutations in BRAF (V600E) with wild-type KRAS and loss of heterozygosity (LOH) of the APC locus.
When comparing somatic analysis tools MuTect (Cibulskis et al., 2013) was superior to the other tools, especially when the tumor allele frequency was low.
DNA was extracted using the Puregene DNA Purification Kit (Gentra Systems, Inc, Wicklow, Ireland) according to manufacturer's instructions from whole blood (germline analysis) or tumor samples (somatic analysis).
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All children undergo a structured diagnostic evaluation consisting of psychological and psychiatric assessment, psychological testing (e.g. intelligence), somatic assessment, analysis of the family structure and/or analysis of videotaped parent child interactions.
A tumour was scored MS-stable when no marker showed instability, MSI-high when > 30% of markers showed instability, and MSI-low when only one marker (<30%) showed instability.[ 22] For somatic mutation analysis of APC, KRAS2, p53 and SMAD4 DNA sequence analysis was performed.
As to be expected for somatic DNA, analysis of Genome 1 DNA yielded no detectable recombinants, confirming that the cloning method for recombinant detection was reliable.
In a second strategy to determine whether cell proliferation is affected in the absence of dRecQ4, normal somatic clone analysis was employed (see Materials and methods for details).
Somatic mutation analysis in NSCLC for molecular staging and the guidance of treatment decisions can be performed on EBUS and EUS fine needle aspirates, procedures that are less invasive for the patient than routine mediastinoscopy.
Informed signed consent for germline and somatic DNA analysis was obtained from each patient recruited by the COMETE network, and the study was formally approved by an institutional review board (CPP Paris-Cochin).
Prior to the evaluation of somatic sets, analysis of GO biological process terms in the GWAS permutation P values rank did not show any significant asymmetry using the Gene Set Enrichment Analysis (GSEA) tool [31] with multiple testing correction by the false discovery rate (FDR) approach [32].
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