OEX014925
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Single-cell multiomics |
Transcriptomic single cell |
The prostates of T2PPRC mice were dissected and minced with scissors. The dissociated prostate cells were resuspended in Red Blood Cell Lysis Solution (Miltenyi Biotec, no. 130-094-183) and incubated at room temperature for 2 min. Dead cells were removed with Dead Cell Removal Kit (Miltenyi Biotec, no. 130-090-101), and the cells were counted via the Countess II FL Automated Cell Counter (Thermo Fisher Scientific). Cells were lysed on ice for 5 min. Isolated nuclei from T2PPRC mouse prostates cells were transposed and loaded onto the Chromium Next GEM Chip J (10X Genomics). Then, the chip with prostate cells was loaded on a Chromium Controller. The library was constructed according to the manufacturer’s instructions. Fifteen mouse prostate samples with different barcodes were run on a NovaSeq6000 (Illumina) (To overcome the limitation of 4.5 months as the lifespan for TPPRC mice due to probable lesions of other organs, we grafted 4.5-month prostate tumors under renal capsule of SCID mice for another 1.5 months to achieve total time of 6 months). |
2021-12-12 |