GSM6821411: Brain, Mock, 12 Months Post Infection [snMultiATAC]; Mus musculus; ATAC-seq
Mice were euthanized via injection with a sublethal dose of ketamine and xylazine followed by exsanguination. Mice were perfused with ice-cold PBS, brain was collected and flash frozen in liquid nitrogen. Brain was processed according to the 10X Genomics Protocol Revision D. Brain was lysed with 0.1x lysis buffer and homogenized wtih scissors and a pellet pestle. Homogenate was incubated for 3 minutes on ice. Suspension was filtered through at 70um filter and run through a 30% Percoll Plus gradient to remove myelin. Cells were then washed, visualized on a microscope to determine nuclear membrane integrity, and counted. Library construction was performed according to 10x Genomics Chromium™ Next GEM Single Cell Multiome ATAC+Gene Expression Reagent Kit protocol single cell gene expression RNA-seq and transposed DNA ATAC-seq