title: single_cell_RNAseq of Mus musculus : adult brain
design: Tissue sections were exposed to 70  of permeabilization enzyme for 18 min, which was selected as the optimal time using the tissue optimization slide (10X Genomics, catalog #1000193), releasing the poly-adenylated mRNA that could then be captured by poly-dT primers containing Illumina TruSeq Read, spatial barcode and UMI on the Visium slides. First and second strand cDNA synthesis and amplification were performed in the Visium slide cassette in a Thermal cycler using a Visium Spatial Gene Expression Reagent Kit (10X Genomics) as per the manufacturers protocol. Dual Indexed, Illumina compatible libraries were created by Sample Index PCR as per the manufacturers instructions using the Dual Index Kit TT Set A (10X Genomics) and E3, E4, E5 and E6 indexing primers. 3 end sequencing was performed in the Princess Margaret Genomics Facility using an Illumina NovaSeq 6000 system.