The genomic DNA was extracted from blood samples from the P1 to P13 patients using a DNeasy blood kit (Qiagen, Valencia, CA) according to the manufacturer’s protocol and then quantified using a Qubit® DNA Assay Kit in a Qubit® 2.0 Fluorometer (Life Technologies). After that, 0.8 μg genomic DNA from each sample with high molecular weight (> 20 Kb single band of DNA after agarose gel electrophoresis) were used for preparation of the DNA library using the TruSeq Nano DNA HT Sample Prep Kit (Illumina, USA) and WGS according to a previous study.32 (link) The raw high-throughput data from P1 to P7 patients were detected with a Illumina Hiseq platform and filtered with the SOAPnuke (v. 1.5) software to remove the adapter sequences and the low-quality reads, whereas the high-quality clean reads were mapped to the human reference genome (hg19; NCBI build GRCh37) with Burrows-Wheeler Aligner (BWA; v. 0.7.12) software.
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Whole Genome Sequencing of Patient Blood Samples
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Variable analysis
independent variables
- DNA extraction method (DNeasy blood kit)
- DNA quantification method (Qubit® DNA Assay Kit)
- DNA library preparation method (TruSeq Nano DNA HT Sample Prep Kit)
- Whole-genome sequencing (WGS) method
- Genomic DNA quantity (quantified using Qubit® 2.0 Fluorometer)
- Genomic DNA quality (high molecular weight > 20 Kb single band after agarose gel electrophoresis)
- Blood samples from P1 to P13 patients
- No positive or negative controls were explicitly mentioned in the provided information.
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