GSM9859774: BRCA 07 PBMC/buffy coat single-cell RNA-seq; Homo sapiens; RNA-Seq
Whole blood was collected into EDTA tubes and processed to obtain the PBMC/buffy coat fraction. Cells were cryopreserved and later thawed for single-cell sequencing. For the 10x-processed ovarian cancer and benign gynecological control samples, thawed cells were transferred with wide-bore tips into warm medium containing 10% FBS, centrifuged, resuspended, and assessed for concentration and viability by trypan blue on a Countess 3 FL counter. Samples with acceptable viability were resuspended in 1x PBS with 0.04% BSA before library preparation. Breast cancer and healthy donor samples were processed from cryopreserved PBMC/buffy coat material by Singleron Biotechnologies. Ovarian cancer and benign gynecological control samples were prepared using the Chromium Next GEM Single Cell 3' Reagent Kit (10x Genomics) following the manufacturer's protocol. Breast cancer and healthy donor samples were prepared using the Singleron GEXSCOPE SCOPE-chip micro-well single-cell RNA-seq system according to the manufacturer's protocol. Libraries were sequenced as paired-end high-throughput sequencing libraries; instrument models are listed in the sample table.