Genome data, epilepsy gene panel, primer for each pathogenic likely pathogenic variant, and Sanger results for Uncovering Etiologic Genes through Whole Exome Sequencing in Pediatric Epilepsy: A Case Series from Thailand

The research project (MTU-EC-PE-1-141/64) was conducted following the Declaration of Helsinki and approved by Thammasat University's Human Research Ethics Committee. We used descriptive cross-sectional statistics to calculate clinically detectable rates. Ten volunteers, aged 0-15, were diagnosed with epilepsy based on specific symptoms and abnormal EEG results. Leukocyte genomic DNA was extracted using a Puregene® blood kit. Exome sequencing was performed utilizing the Illumina HiSeq platform (Macrogen, South Korea). All variant annotations were examined using a standard methodology (the Burrows-Wheeler Alignment tool [BWA]; http://bio-bwa.sourceforge.net/bwa.shtml) . The variants were then filtered using a minor allele frequency (MAF) of >0.01 in the database for single nucleotide polymorphisms in the entire 1000 genome data (phase 3). The gene target analysis approach was applied for phenotypes based on the Human Phenotype Ontology (HPO) keywords of epilepsy. (HP:0001250). The gene list is included in the data availability. The categorization for genetic variant interpretation was based on guidelines from the American College of Medical Genetics and Genomics and the Association for Molecular Pathology's variant classification criteria. Franklin (https://franklin.genoox.com), Clinvar (https://www.ncbi.nlm.nih.gov/clinvar), gnomAD (https://gnomad.broadinstitute.org), and our Thai database (https://trex.nbt.or.th/home) were used as in silico predictive programs to determine the novelty of variants and the supposed evidence of pathogenicity of identified variants. Sanger sequencing was used to confirm the existence of pathogenic and potentially pathogenic mutations in both patients and parents. A polymerase chain reaction was utilized. 10 mMol of dNTP-Mix (GeneON®, lot number 101.183) volume 0.5 microliter (uL) with 10xTaq buffer (Thermo Scientific®, lot number 00892578) Volume: 2.5 uL, 5 uM/uL of forward and reverse primer (Macrogen®) Volume 1.5 uL per each (sequence of forward and reverse in Supplementary tablexxx), 25 mM MgCl2 (Thermo Scientific, lot number 00892580) Volume: 1.5 uL, Taq DNA Polymerase Recombination (Thermo Scientific, lot number 2667134) Volume: 0.2 uL, DNA (50 ng/uL), 3 uL, and up to 25 uL of water per reaction.