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Study of microglia in aging and in a tissue specific manner (white and grey matter) using Smart-seq2 and 10x scRNA-seq technologies. We also analysed, 5xFAD mice (AD model) with Smart-seq2, and Apoe-KO with 10x transcriptomics. Additionally we studied the impact of Actinomycin-D during tissue dissociation on gene expression in microglia using Smart-seq2.

Grey and white matter (corpus callosum) were dissected from the brain of 4 months and 21 months old wild type mice, single cell suspension was isolated using Neural Tissue Dissection kit (myelin was removed on percoll gradient) then microglia cells were isolated using CD11b microbeads by Magnetic separation with MS columns (MACS technology).The cell pellets are resuspended in the RLT buffer (100ul-150ul), snap frozen in liquid nitrogen and stored in -80 freezer. RNA was extracted and transcriptome ...

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