GSM9314724: Single-cell RNA sequencing of mouse tissue with BiTE EVs treated, A2; Mus musculus; RNA-Seq
Fresh tissues were preserved in sCelLiveTM Tissue Preservation Solution (Singleron) on ice within 30 min post-surgery. Specimens were washed with HBSS, minced, and digested with 3 mL sCelLiveTM Dissociation Solution (Singleron) (37°C, 15 min) using Singleron PythoN™ Tissue Dissociation System. After filtration (40 μm strainer), red blood cells were lysed with GEXSCOPE® RCLB (1:2 ratio, 5-8 min, RT). The mixture was then centrifuged at 300 × g 4 ℃ for 5 mins to remove supernatant, followed by PBS resuspension. Cell viability was assessed via Trypan Blue staining Single-cell suspensions (2×105 cells/mL in PBS) were loaded onto a microwell chip using the Singleron Matrix® Single Cell Processing System. Barcoding Beads were collected from the chip, and reverse transcription was performed to generate cDNA from captured mRNA, followed by PCR amplification. The amplified cDNA was fragmented, ligated with sequencing adapters, and libraries were constructed using GEXSCOPE® Single Cell RNA Library Kits (Singleron)