GSM8061827: breast cancer tissue, patient 931881; Homo sapiens; RNA-Seq
Fresh breast cancer tissues and the corresponding normal tissues were stored in the sCelLive® Tissue Preservation Solution (Singleron Biotechnologies, Nanjing, China) on ice immediately after surgery. Tissues were then washed with Hanks Balanced Salt Solution (HBSS) for 3 times, minced into small fragments, and digested in sCelLive® Tissue Dissociation Solution (Singleron Biotechnologies) at 37 °C for 15 minutes. The dissociated tissues were filtered through 40 μm cell strainers. The red blood cells were removed by GEXSCOPE® red blood cell lysis buﬀer (Singleron Biotechnologies) at room temperature for 6 minutes. The single cell suspensions in PBS were loaded into microfluidic devices. Single-cell suspensions (2×105 cells/mL) with PBS (HyClone) were loaded onto microwell chip using the Singleron Matrix®Single Cell Processing System. Barcoding Beads are subsequently collected from the microwell chip, followed by reverse transcription of the mRNA captured by the Barcoding Beads and to obtain cDNA, and PCR amplification. The amplified cDNA is then fragmented and ligated with sequencing adapters. The scRNA-seq libraries were constructed according to the protocol of the GEXSCOPE® Single Cell RNA Library Kits (Singleron).