Exome sequencing for genetic diagnosis
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Corresponding Organization :
Other organizations : Stanford University, Smith-Kettlewell Eye Research Institute, University of Iowa, Palo Alto University, National Institutes of Health, National Eye Institute, New York University, Columbia University, Palo Alto Veterans Institute for Research
Variable analysis
- Genomic DNA from the proband and parents were processed using the Agilent Clinical Research Exome kit
- The targeted regions (exonic regions and flanking splicing junctions) were sequenced simultaneously by massively parallel (NextGen) sequencing on an Illumina HiSeq sequencing system with 100-bp paired-end reads
- Sequence variants identified in the proband and parent samples
- Mean depth of coverage of 104× (quality threshold 95.3%)
- Bidirectional sequences were assembled, aligned to reference gene sequences based on GRCh37/UCSC hg19
- Variants were filtered based on predicted loss-of-function or missense mutation, presence of gene or variant in the Human Gene Mutation Database, and MAF of <0.01
- Capillary sequencing was used to confirm all potentially pathogenic variants identified in the proband and parent samples
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