WGS of HGSOC: microdissected region P1.background - 1
Frozen tissue was sectioned (10 µm) and mounted to standard superfrost slides, methanol fixed, stained with hematoxylin and eosin, and scanned on a CellCelector (ALS). Using the CellCelector, the long edge of a 150 µm glass capillary was used to mechanically scrape small tissue sections from the slide which were aspirated and deposited in 1 µL of PBS (10 mM Phosphate, 2.68 mM Potassium Chloride, 140 mM Sodium Chloride, 18912014, Thermo Fisher Scientific) in 0.2 mL PCR tubes (Eppendorf). Tissue sections were subjected to whole genome amplification using the Ampli1 WGA Kit (Silicon Biosystems) according to the manufacturer's instructions. Following amplification, 400 bp sequencing libraries were constructed using the Ampli1 Low-Pass Whole Genome Sequencing Kit for Ion Torrent (Silicon Biosystems) according to the manufacturer's instructions. Libraries were diluted to 50 pM, loaded into an Ion Chef for template preparation and loading into an Ion 530 chip, and then sequenced for 525 flows on an Ion S5 (Thermo Fisher Scientific).