GSM7034566
Anti-mesothelin M5 CAR-TCONV and CAR-TRM cells were harvested after nine days of in vitro expansion. CD8+ CAR T-cells were magnetically isolated using anti-mesothelin-coated beads (ACROBiosystems) and anti-CD8 microbeads (Miltenyi Biotec). After elimination of dead cells, 100,000 CD8+ CAR T-cells were cryopreserved. Library preparation was performed by Novogene as follows: a reaction mixture consisting of Tn5 transposase and equimolar adapter1 and adapter 2 was added to nuclei pellets and incubated at 37°C for thirty minutes. The library was enriched using PCR, and purification of final libraries was carried out using AMPure beads (Beckman Coulter). Paired-end sequencing (150bp reads) was conducted on a NovaSeq 6000 instrument.