Blood samples obtained from the proband were sent to the Nanfang Hospital Precision Medicine Center for Whole-exome sequencing (WES) of genomic DNA. Illumina HiSeq platform was used for WES. The criteria established and revised by the American College of Medical Genetics and Genomics (Richards et al., 2015 (link)) was used to classified the variants. Two hypertriglyceridemia associated genes mutations, within exon 10 of the LMF1 gene and exon 5 of the LPL gene identified by WES, were verified using Sanger sequencing. Standard phenol/chloroform extraction was performed to extract genomic DNA from the peripheral blood acquired from the proband and his family members (Ⅰ1, Ⅰ2, Ⅱ4, Ⅱ2, Ⅲ4, and Ⅲ3). The PCR primers that were designed using Primer-BLAST (https://www.ncbi.nlm.nih.gov/tools/primer-blast ) were as follows: LMF1: Exon-10 forward primer: 5′-CCGTCTCAGCCACCAGAAAA-3′, Exon-10 reverse primer: 5′-CACGGCTGGTTTGGTTTGAG-3'; and LPL: Exon-5 forward primer: 5′-CCAGCCATCCTGAGTGGAAA-3′, Exon-5 reverse primer: 5′-GGCTCTAAGGTGGTCATGCT-3'.The PCR products were then analyzed by agarose gel electrophoresis and submitted to Invitrogen (Shanghai, China) for Sanger sequencing.
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