MAPseq and BARseq Dual Sectioning Event for a Single Mouse Brain v1

This protocol enables tissue processing for combined BARseq- MAPseq experiments where axonal projections of individual neurons transduced by unique barcode sequences can be mapped across the brain, with concurrent molecular characterization of transduced cells by gene expression. Mice receive injections of Sindbis virus barcode library targeting region of interest, and fresh brain tissue will be harvested and frozen in OCT 24 hours later. Frozen brain tissue will be stored at -80°C to preserve RNA until ready for sectioning and subsequent processing for in situ sequencing of genes and barcodes and next generation sequencing projection barcodes in dissected tissue.