GSM8566271: basal CK5PIP, 2m, mouse 2, plate 1; Mus musculus; RNA-Seq
Whole mouse prostates were harvested and digested to single cell solution using 5mg/mL collagenase in advanced DMEM (+/+/+) for 1 hour of shaking at 37 degrees Celsius. This was followed by medium washes and 15-minute incubation with TrypLE at 37 degrees Celsius then the digest solution was passed through a 70uM filter. Cells treated with DAPI for live/dead stain were sorted at the Department of BioMedical Research FACs Core facilities in a solution of PBS with 0.2% fetal bovine serum. Protocols for sorting single eYFP+ cells into wells of 384-well SORT-Seq plates were conducted under the recommendations of Single Cell Discoveries (https://www.scdiscoveries.com/). eYFP positive control prostates were used to adjust A488 gating settings for each sort. Two animals and three 96-well plates were collected as single cells for each group, centrifuged at 4 degrees Celsius, snap-frozen on dry ice, and then stored at -80 degrees Celsius until transport to sequencing facilities The SORT-seq protocol (a partially robotized method based on the CEL-Seq2 protocol) was used to generate libraries for scRNA-Seq