Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Mice were sacrificed by an overdose of Isoflurane or Ketamin/Xylazine, followed by perfusion through the left ventricle with artificial cerebrospinal fluid (aCSF), equilibrated in 95%O2/5%CO2 on ice or 4°C. The dentate gyrus was then microdissected from 300 μm vibratome brain sections. Single-cell suspensions were prepared using the Papain kit (Worthington) with 25-35min enzymatic digestion followed by manual trituration using a BSA-coated p1000 pipette or fire-polished Pasteur pipettes. For hGFAP-GFP mice, GFP-positive cells were sorted on a BD FACSAria II into oxygenated aCSF at 4°C. 1000 cells/ μl were resuspended in artificial cerebrospinal fluid and added to the Reverse transcription mix to aim for sampling of around 3500 cells. All downstream cDNA synthesis (13 PCR cycles), library preparation and sequencing were carried out as instructed by the manufacturer (10X Genomics Chromium Single Cell Kit Version 2).