Mouse_Brain__Cortex__Single_Nuclei_Sequencing_Protocol_Pilot_Study

Fresh frozen mouse brain was sectioned on a cryostat, followed by individual microdissections of mouse brain cortical areas. Tissue was homogenized on the same day using dounce homogenizer, then filtered and FACS stained for nuclei sorting. Finally, FACS sorted individual nuclei were used for 10x RNA sequencing library construction. Cellular morphology of brain cells, such as neurones, make it extremely difficult to purify these populations without causing damage to their structure, and subsequently, transcriptome. Single nuclei sequencing has been proven to be a good alternative to standard single cell sequencing and capable of replicating the same expression signature as single cells. This is a pilot study for mouse brain single nuclei sequencing. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/