Xt inherited disease panel cat no 5190 7519

Patients’ genomic DNA was isolated from 2‐ml peripheral blood samples using a QIAamp Blood DNA Mini Kit® (Qiagen GmbH, Hilden, Germany). Three micrograms of DNA was processed through shearing using a Covarias® M220 Ultrasonicator system (Covaris, Inc. Woburn, MA, USA) to pieces of 150–200 bp in size. An adapter‐ligated library was produced with Agilent SureSelect Target Enrichment System (Agilent Technologies, Inc., Santa Clara, CA, USA) according to the manufacturer's instructions. The capture library was performed using an XT Inherited Disease Panel (cat No. 5190–7519, Agilent technologies, Inc.), containing 2,742 genes known to cause inherited disorders, covering only 10.5 Mb. Clusters were then generated by isothermal bridge amplification using an Illumina cBot station, and sequencing was performed on an Illumina HiSeq 2500 System (Illumina, Inc., San Diego, CA, USA). The raw data (fastq file) for each patient were obtained for CNV identification. The average sequencing depth of data used was 122, and more that 95.6% of targeted region was covered with 20 reads.

The patient's and her parents' genomic DNA were extracted from 2-ml peripheral blood samples using the QIAamp DNA Blood Mini Kit (Qiagen GmbH, Hilden, Germany). The DNA concentration and purity were measured using an Invitrogen Qubit dsDNA detection kit and a Qubit4 fluorometer (Carlsbad, CA, USA). The Agilent Sure Select Target Enrichment System (Agilent Technologies Inc., Santa Clara, CA, US) was used to produce an adapter-ligated library according to the manufacturer's instructions. An XT Inherited Disease Panel (cat No. 5190–7519, Agilent Technologies Inc.) containing 2,742 genes was used to create the capture library. The clusters were then produced using an Illumina cBot Station and sequenced on an Illumina HiSeq 2,500 System (Illumina Inc., San Diego, CA, US). Using human genome hg19 as the reference, alignment of sequence and repeated labeling were performed using BWM version 0.7.17 (http://bio-bwa.sourceforge.net/) and Picard bioinformatics software version 2.5.0 (https://broadinstitute.github.io/picard/) for biological analysis and interpretation. GATK 4.0.0.0 (https://gatk.broadinstitute.org/hc/en-us/sections/360007407851-4-0-0-0) and Samtools 1.8 (https://sourceforge.net/projects/samtools/files/samtools/1.8/) were used to identify mutation sites.