GSM9512691: SCLC_JSJ; Homo sapiens; RNA-Seq
Tissues were placed in MACS Tissue Storage Solution (Miltenyi Biotec) until processing. The BD Rhapsody system was used to capture the transcriptomic information of the sample-derived single cells. Single-cell capture was achieved by random distribution of a single-cell suspension across > 200,000 microwells by a limitingdilution approach. Beads with oligonucleotide barcodes were added to saturation so that a bead was paired with each cell. The cells were lysed in the microwell to hybridize mRNA molecules to barcoded capture oligos on the beads. Beads were collected in a single tube for reverse transcription and ExoI digestion. Each cDNA was tagged at the 5′-end (i.e., the 3′-end of an mRNA transcript) with a unique molecular identifier (UMI) and cell barcode indicating its cell of origin. Whole-transcriptome libraries were prepared using the BD Rhapsody single-cell whole-transcriptome amplification (WTA) workflow, including random priming and extension (RPE), RPE amplification PCR, and WTA index PCR. The libraries were quantified using a High-Sensitivity DNA Chip (Agilent) on a Bioanalyzer 2200 and the Qubit High-Sensitivity DNA assay (Thermo Fisher Scientific). Sequencing was performed using an Illumina sequencer (San Diego, CA, USA) on a 150-bp paired-end run.