GSM9158969: SKBR3- riGFP-HER2 CAR T, Mouse 1; Homo sapiens; RNA-Seq
Tumors were excised and enzymatically digested with Liberase TM (75 µg/mL) and DNase I (20 µg/mL) at 37°C for 40 minutes, followed by mechanical dissociation and filtration through a 70 µm strainer. Single-cell suspensions were stained for viability and surface markers. SKBR3 tumor cells were sorted by FACS based on positive staining for human EpCAM and exclusion of mouse cells (CD45⁻, CD31⁻, Podoplanin⁻). Viable tumor cells were sorted and RNA was extracted using the Qiagen RNeasy Micro Kit following the manufacturer's protocol RNA quality was assessed using an Agilent Bioanalyzer. Strand-specific mRNA-seq libraries were prepared using the TruSeq mRNA-SEQ Library Preparation Kit (Illumina), following the manufacturer's instructions. Libraries were quantified and sequenced using the Illumina NovaSeq 6000 platform.