P08_Dura_Tapestri
Nucleus were isolated from cryopreserved tumour and meningeal tissue samples from patients P01 and P02. Tissues (50100 mg) were minced on ice and homogenised in 1 mL of lysis buffer (comprising 0.2% Triton X-100, 1  Roche protease inhibitor, 1 mM DTT, and 0.2 U/L RNAsin in 2% BSA in PBS). The nuclei pellet, recovered via centrifugation at 500g for 5 minutes at 4C, was resuspended in 400 L of suspension buffer (1 mM EDTA, 0.2 U/L RNAsin, and 2% BSA in PBS). After filtration through a 40m strainer and DAPI staining (0.5 g/mL), DAPI-positive nuclei were isolated using a flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) following a secondary 20m filtration step. A custom multiplex amplicon panel was designed via Tapestri Designer (MissionBio), incorporating exons of NF2, TRAF7, KLF4, and AKT1, alongside target regions on chromosome 22q, into the standard Tapestri glioma panel. This final configuration comprised 297 amplicons and 1,272 targets (total size 52.9 kb). Approximately 100,000 nuclei per sample were loaded onto a Tapestri microfluidic cartridge for encapsulation with lysis reagents, followed by protease inactivation. Within the droplets, cell-identifying molecular barcodes were attached to targeted genomic regions using gene-specific primers and hydrogel beads. After UV-mediated primer cleavage and targeted PCR amplification, the emulsions were broken and the DNA was purified using AMPure XP beads (Beckman Coulter, Brea, CA, USA). Following the addition of Illumina adapter sequences via a 10-cycle PCR and a final purification, library quality was verified on a 4200 TapeStation (Agilent Technologies, Santa Clara, CA, USA). Sequencing was performed on an Illumina NovaSeq 6000 system (Illumina, San Diego, CA, USA) with 2$\times$150 bp paired-end reads. Experimental procedures from library preparation to sequencing were conducted by JSLink, Inc. (Seoul, Republic of Korea).