Fig. 1

Typical workflow of NGS and bioinformatic analysis. Genomic DNA is extracted and massive parallel sequencing on various NGS platforms is performed. For whole exome sequencing, exomes capture is performed, and the raw sequences are aligned to reference genome. Variant calling is performed and filtered by various softwares. Common variants can be further analyzed for risk assessment in common diseases. Non-synonymous and rare variants can be further scrutinized according to the presumed inheritance pattern and deleteriousness predictions. Candidate variants can be confirmed by validation, segregation analyses and pathogenicity studies. For novel variants, additional affected patients and families with the same variant can further strengthen the association of the variant with disease