title: GSM8410132: Qpct-knock out,replicate 1,snRNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: The nuclei of E15.5 mouse placenta (Qpct+/+ and Qpct-/-) were isolated as described in Marsh and Blelloch (2020). The placental tissue was cut into small pieces (<1mm) and placed into 2mL Nuclei EZ lysis buffer (Sigma, St. Louis, MO-NUC-101). The tissue was transferred to a dounce homogenizer with an additional 2mL Nuclei EZ lysis buffer. The nuclei were homogenized 10 times with a loose pestle (A) and 10 times with a tight pestle (B). The nuclei solution was transferred to a 15mL Falcon tube, and 2mL Nuclei EZ lysis buffer was added and incubated on ice for 5 minutes. The solution was passed through a 40μm filter and centrifuged at 500g for 8 minutes at 4℃. The supernatant was discarded, and the pellet was resuspended in 2mL Nuclei EZ lysis buffer, incubated on ice for 5 minutes, and centrifuged again at 500g for 5 minutes at 4℃. The supernatant was removed, and 2mL Nuclei Wash and Resuspension Buffer was added without mixing and incubated on ice for 5 minutes. This step was repeated once. The final nuclei solution was passed through a 35μm filter. The isolated nuclei were loaded into a Chromium Single Cell 3' Library & Gel Beads kit v3 i7 Multiplex kit (10× Genomics).