title: GSM8948194: CTCs isolated from patient 021, sample 8; Homo sapiens; RNA-Seq
library_name: GSM8948194
library_construction_protocol: Blood samples were obtained via blood draw from multiple locations (via port, left brachiocephalic, superior vena cava, right atrium, portal vein, inferior vena cava, right iliac vein, left common iliac vein, aorta) in ACD-A vacutainers (Fisher Scientific, BD364606) and supplemented with 0.5 µg Tirofiban (Sigma-Aldrich, 30165) per ml of blood. Upon arrival, the samples were equilibrated at room temperature for 15 minutes. Subsequently, 7.5 µl of 0.5 M EDTA (Invitrogen, AM9260G) per ml of blood was added. The samples were then processed for microfluidic-based CTC capture within approximately 48 hours of the blood draw. Using the Parsortix Cell Separation System (ANGLE), CTCs were captured in Cell Separation cassettes (GEN3D6.5) and stained with an antibody cocktail containing anti-human EpCAM–AF488 (Cell Signaling Technology, CST5198), HER2–AF488 (BioLegend, 324410), EGFR–FITC (GeneTex, GTX11400), and CD45-AF647 (BioLegend, 304018).  The cDNA was processed using FLASH-seq protocol. After enrichment PCR, the samples were cleaned up with 0.8X AMPure Beads XP (Beckman Coulter, A63881) and the cDNA concentration was measured with the Tapestation and a High Sensitivity D5000 Screen Tape (Agilent). Libraries were prepared using Nextera XT (Illumina) and sequenced on an Illumina NovaSeq X Plus instrumen.
identifiers.namespace: GEO
identifiers.id: GSM8948194_r1