GSM7863753: C42B Xenograft ENZ 3W; Homo sapiens; RNA-Seq
C4-2B xenograft and TRAMP tumor tissues were harvested and dissociated using “Tumor Dissociation Kit, human, #130-095-929” and “Tumor Dissociation Kit, mouse, #130-096-730” from Miltenyi Biotec. Samples were assessed and processed for single cell 3' sequencing. GEMs (Gel Bead-in Emulsions) were generated in a Chromium Controller (10x Genomics). RNA contained in the GEMs was reverse transcribed, barcoded, and cDNA was recovered. cDNA was assessed with the Agilent TapeStation D5000 High Sensitivity Kit (Agilent Technologies). All samples underwent manual library processing with the Chromium Next GEM Single Cell 3' Reagent Kit v3.1: cDNA was fragmented, processed, and size selected. Adaptors were ligated and samples were cleaned up. Dual indexes were attached via PCR. The completed libraries were cleaned up and assessed for quality with the Agilent TapeStation D5000 High Sensitivity Kit. Libraries were quantitated with qPCR (NEBNext Library Quant Kit for Illumina, Bio-rad CFX96 Touch Real-Time PCR). Libraries were normalized to 1.5 pM and pooled. The library pool was denatured and diluted to approximately 300 pM. A library of 1.5 pM PhiX was spiked in as an internal control