title: GSM8793205: Astrocyte mCherry OE, replicate 2, scRNAseq; Mus musculus; RNA-Seq
library_name: GSM8793205
library_construction_protocol: P15 mice were perfused with 15 mM HEPES pH7 + 0.5% Glucose + 1xHBSS supplemented with 5ug/mL Actinomycin D and 10 uM Triptolide. Brains were extracted and cortices were dissected off in ice cold 1X HBSS supplemented with 5ug/mL Actinomycin D, 10 uM Triptolide, and 27.1 ug/mL Anisomycin. Cortices were then dissociated using the MIltenyi GentleMACS system in Miltenyi Neural Tissue Dissociation kit supplemented with 5ug/mL Actinomycin D, 10 uM Triptolide, and 27.1 ug/mL Anisomycin. Enzymes were washed off and single cell suspension was generated by passing cells through a 70 um cell strainer. Single cell suspension was then resuspended in MACS buffer and cells were incubated with CD11b selection beads. Excess beads were washed off with MACS buffer and cells were then passed through a 30 um cell strainer into LS columns. CD11b+ fraction was collected and cells were resuspended in Cell suspension buffer following PIPseq kit instructions. Up to 40,000 cells were added to PIP reaction and processed for library preparation as instructed by the PIPseq kit. Library construction was performed following manufacturer's instructions from PIPseq v4.0PLUS kit.