Our custom hybrid capture panel covers coding exons and regions prone to mutations of 10,929 genes that had variants in the above WES results or had been reported in the literature of EC [6 (link),7 (link),8 (link),19 (link)]. The list of targeted genes is in Table S2 . Oligonucleotide probes were designed by BGI. The probe regions were 4.96 Mb, and the average sequencing depth was 20,000×.
All the samples were profiled by targeted panel sequencing (TPS). Fresh surgical samples (tumor and normal) and PIP-E samples were stored at −80 °C until DNA extraction. PAP-C and SWAB-V samples were preserved in ThinPrep PreservCyt solution at 4 °C (Hologic, Marlborough, MA, USA). After delivery to the laboratory, PAP-C and SWAB-V samples were centrifuged for 5 min at 14,000× g, and the precipitates were used for DNA extraction. DNA extraction, library preparation, and targeted NGS were performed as described above. Raw sequencing data of EC patients were also processed like WES data. Sequencing data from women with risk factors for EC were identified as variants using the HaplotypeCaller module of GATK.
All the samples were profiled by targeted panel sequencing (TPS). Fresh surgical samples (tumor and normal) and PIP-E samples were stored at −80 °C until DNA extraction. PAP-C and SWAB-V samples were preserved in ThinPrep PreservCyt solution at 4 °C (Hologic, Marlborough, MA, USA). After delivery to the laboratory, PAP-C and SWAB-V samples were centrifuged for 5 min at 14,000× g, and the precipitates were used for DNA extraction. DNA extraction, library preparation, and targeted NGS were performed as described above. Raw sequencing data of EC patients were also processed like WES data. Sequencing data from women with risk factors for EC were identified as variants using the HaplotypeCaller module of GATK.
Free full text: Click here
