GSM9759714: Intestinal organodis, uninfected; Homo sapiens; RNA-Seq
The human intestinal organoids was suspensed in the digestion buffere TrypLE for 30 min for reaching single cell state. The cells were resuspended in DMEM/F12 and spun at 500 g for 5 min at RT. The supernatant was removed by the pipet, and the cell suspensions were resuspended in PBS containing 0.04% BSA. The suspensions were passed through a 30 µm cell strainer and then washed with PBS containing 0.04% BSA 3 times. Resulting cell suspensions were used for a single-cell RNA sequencing kit from 10xGenomics Single-cell suspensions were loaded onto the 10x Chromium controller using the 10xGeomics Single Cell 3' Library Kit v.2 (10xGenomics) and Chromium Next GEM Single Cell 3' Reagent Kits v.3.1 (10xGenomics) following the manufacturer's guideline. Briefly, single cells and beads were generated, followed by reverse transcription, cDNA amplification, fragmentation, and ligation with adapters with unique sample index PCR. Constructed libraries were sequenced with Illumina NovaSeq by Novogene