Seqcap ez prime choice library kit

Genotyping of 115 variants in ESSE-Ivanovo and RPS was performed by NGS. The libraries for the NGS custom panel were prepared using the SeqCap EZ Prime Choice Library kit (Roche, Basel, Switzerland). Exome libraries were prepared using IDT-Illumina TruSeq DNA Exome protocol (Illumina, San Diego, CA, USA). Sequencing was performed on a Nextseq 550 (Illumina, San Diego, CA, USA). All sequencing stages were performed according to the manufacturers’ protocols.

Genomic DNA was isolated from peripheral blood with the QIAamp DNA Blood Mini Kit (Qiagen, Hilden, Germany). DNA concentration was measured on the Qubit 4.0 fluorimeter (Thermo Fisher Scientific, Waltham, MA, USA). For next-generation sequencing (NGS), the libraries were prepared using either the SeqCap EZ Prime Choice Library kit (Roche, Basel, Switzerland) for targeted sequencing (custom panel) or the IDT-Illumina TruSeq DNA Exome protocol (Illumina, San Diego, CA, USA) for exome libraries. The custom panel consisted of 25 genes (CDS + 25 bp padding), associated with lipid metabolism disorders, including APOE (ABCA1, ABCG5, ABCG8, ANGPTL3, APOA1, APOA5, APOB, APOC2, APOC3, APOE, CETP, GPD1, GPIHBP1, LCAT, LDLR, LDLRAP1, LIPC, LIPI, LMF1, LPL, MTTP, PCSK9, SAR1B, STAP1, USF1). Sequencing was performed on a Nextseq 550 (Illumina, San Diego, CA, USA). All stages of sequencing were conducted according to the manufacturers’ protocols.