GSM4116571: Quadriceps; Mus musculus; RNA-Seq
Each tissue was minced in 2.5 ml digestion solution (1 U/ml collagenase B and 1 U/ml dispase II (Roche Diagnostics, Indianapolis, IN, USA) in PBS) and incubated at 37ºC for 1 hour. The reaction was terminated by adding 10 ml PBS containing 10% fetal bovine serum (FBS). The mixture was then filtered through a 70-m cell strainer and centrifuged at 250 g for 5 minutes. The pellet was collected and the supernatant was centrifuged again at 250 g for 5 minutes. The pellet was combined with the pellet from the first centrifugation, washed with PBS, and centrifuged at 670 g for 10 minutes. The pellet was re-suspended in 3 ml PBS and filtered through a 40-m cell strainer. For brain, cell suspension was layered on equal volume of 50% isotonic Percoll (Sigma-Aldrich) and centrifuged at 600 g for 25 minutes. Supernatant was then discarded and cell pellet was re-suspended in FACS staining buffer.The cell suspension was layered on equal volume of the Lympholyte-M solution (Cedarlane, Burlington, NC, USA) and centrifuged at 2,095 g for 45 minutes. The library was generated according to manufacturer's instructions (10X Genomics). Expression profiling by high throughput sequencing
GEO Accession
GSM4116571