Ten microlitres of each normalised DNA library were pooled and incubated at 96 °C for 2 min. The library pool solution was mixed, centrifuged briefly and incubated on ice for 5 min. Libraries were sequenced on an Illumina Nova-Seq instrument. DNA libraries were prepared using the hybrid capture-based TruSight Oncology 500 (TSO500) Library Preparation Kit (Illumina, San Diego, CA, USA) following Illumina’s TSO500 reference guide targeting 523 cancer-relevant genes with a target capture size of 1.94 megabases (Mb) (see supplementary data 2 for a list of genes). The panel includes the following Meningioma-related genes and targets from the literature: NF2, KDM5C, KDM6A, SMARCB1, AKT1, mTOR, SMO, TRAF7, KLF4, PIK3CA, BAP1, TP53, CDKN2A, CDKN2B, TERT (+ promoter), NAB2-STAT6 fusion. Mean sequencing yield in the TSO500 data was 18 gigabases (Gb) per sample, with a mean unfiltered depth of coverage of 9278 reads per sample.
A subset of 13 tumours was also sequenced using the Agilent SureSelect Clinical Research Exome V2 (Agilent, Santa Clara, CA, USA) with a capture size of 67.3 Mb. Mean sequencing yield in the exome data was 46 Gb per sample, with a mean unfiltered depth of coverage of 688 reads per sample.
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