GSM2804357: WT 1xLPS7; Mus musculus; RNA-Seq
10k microglia were directly sorted (using a Sony SH800 cell sorter) into RNAse-free PCR strips containing 30 μl of H2O with 0.2% Triton-X and 0.8 U/μl RNAse inhibitor (Clontech) and samples were immediatedly frozen on dry ice. . RNA was isolated using NucleoSpin RNA XS kit (Macherey-Nagel) according to the manufacturer’s instructions. 3ng total RNA was used as input material for cDNA synthesis. cDNA synthesis and enrichment was performed following the Smart-seq2 v4 protocol as described by the manufacturers (Clontech). Sequencing Library was prepared with 1 ng of cDNA using the Nextera XT library preparation kit (Illumina) as described59. Multiplexing of samples was achieved using 3 different index-primers in each lane. For sequencing, samples from each group (APP and WT) were pooled to rule out amplification and sequencing biases.