title: GSM7163470: Kidney organoids generated from 0.5K cells; Homo sapiens; RNA-Seq
library_construction_protocol: After the indicated time point kidney organoids were collected and washed twice with PBS, and further dissociated into a single cell suspension by treating them with Accumax (07921, Stem Cell Technologies) for 15 min at 37ºC followed by Trypsin-EDTA 0.25% (wt/vol) trypsin (25300-054, Life Technologies) for additional 15 min at 37ºC. The reaction was deactivated by adding 10% FBS. The cell suspension was then passed through a 40μm cell strainer. After centrifugation at 1,000 RPM for 5 minutes cell numbers and viability were analyzed using Countess Automated Cell Counter (Invitrogen). This method generated single cell suspension with greater than 90% viability. Cell suspensions were frozen until further use. For library construction cells were thawed and centrifuged at 1,000 RPM for 5 minutes. Cell numbers and viability were analyzed using Countess Automated Cell Counter (Invitrogen) and cell suspensions were loaded onto a well of a 10X Chromium Single Cell instrument. Barcoding and cDNA synthesis were performed according to the manufacturer´s instructions. The cDNA libraries were constructed using the 10X Chromium Single cell 3' Library Kit v3 according to the manufacturer's original protocol and sequenced with a NextSeq 500 (R1:28, R2:55, i7:8) instrument. scRNA libraries were prepared for sequencing using standard Illumina protocols