NGS of homo sapiens: adult female blood
DNA fragments, containing all coding exons and exon-intron junctions, were enriched by solution-based hybridization capture, followed by sequencing using the Illumina HiSeq 2000 platform (Illumina, Inc., San Diego, CA, USA). Analysis of next-generation sequencing data was performed using an in-house analysis pipeline. Briefly, sequencing reads from the HiSeq 2000 raw data were sorted by index and barcode sequences. Sorted fastq files were aligned to the hg19 reference genome using the Burrows-Wheeler Aligner (BWA; ver. 0.7.12) BWA-MEM algorithm. Output SAM files were converted into BAM files and sorted using SAMtools (ver. 1.1). Duplicate removal was performed with Picard tools (ver. 1.128) MarkDuplicates. Realignment around known indel sites and Base Quality Score Recalibration (BQSR) were performed using GATK (v3.3.0) to create the final BAM files. Variants were called using the GATK v3.3.0 Unified Genotyper algorithm for loci with sequencing depth greater than or equal to 50X.