title: GSM6940186: CNS endothelial cells, ETX, 1; Mus musculus; RNA-Seq
library_name: GSM6940186
library_construction_protocol: CNS tissue was enzymatically dissociated with a papain solution followed by vigorous trituration and a second dissociation with collangese and dispase solution.  Myelin was removed using Miltenyi Biotec Myelin Removal Beads II per the manufacturer's instructions.  Isolated cells were stained with anti-CD31 clone 390, anti-CD45 clone 30-F11 (, and CD11b clone M1/70, anti-CD13 clone R3-242, anti-PDGFbeta clone APB%, and DAPI.  Viable endotheilail cells (DAPI-) positive for CD31 only (CD31+, CD45-, CD11b-, CD13-, and PDGFbeta -) were sorted via FACS using a BD Biosciences FACSAria II Cell Sorter.  RNA was extracted from sorted endothelial cells using Qiagen's RNeasy Plus Micro Kit per the manufacturer's instructions. cDNA was generated using Takara's SMART-Seq v4 Ultra Low Input RNA Kit. Total RNA integrity was checked using a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). The cDNA synthesis and amplification were performed by SMART-Seq v4 ultra low input RNA kit (Takara Bio USA, Mountain View, CA, USA) starting with less than 1 ng of total RNA from each sample. 