NGS OF HUMAN SAMPLE
Genomic DNA of BC, OC and PC patients were collected using buccal swabs and extracted through MagPurix instrument and Forensic DNA Extraction Kit (Zinexts Life Science Corp.- CodZP01001) according to the manufacturer's protocol. NGS was executed by the Ion Torrent S5 system (Thermo Fisher Scientific, Waltham, MA, USA) after automatic library preparation using Ion Chef (Thermo Fisher Scientific, Waltham, MA, USA). Ion Chef consists of fragmentation and adapter ligation onto the PCR products, clonal amplification. The DNA libraries were quantified with Real-Time Step One PCR System (Thermo Fisher Scientific, Waltham, MA, USA) and the prepared samples were loaded onto an Ion 530 chip by Ion Chef (Thermo Fisher Scientific, Waltham, MA, USA). Ion S5 Plus (Thermo Fisher Scientific, Waltham, MA, USA) instrument was used for the sequencing. Specific plugins as SampleId and Coverageanalysis were used for NGS data analysis on the Torrent Suite 5.14.0 platform. The uniformity of base coverage was over 98% in all batches, and base coverage was over 20 at all target regions. This NGS method cannot detect variations outside the +/-10 nucleotide coding sequence.