Dataset features


Application: RNA-seq analysis
Number of samples: 19
Release date: Aug 24 2011
Last update date: Jul 20 2018
Access: Public
Diseases: Adrenal Cortex Diseases
Dataset link A Transcriptomic Atlas of Mouse Neocortical Layers

Experimental Protocol

Eight adult male mice (56 days old; C57BL/6J strain) were killed by cervical dislocation according to approved schedule one UK Home Office guidelines (Scientific Procedures Act, 1986). The eight were comprised of two groups of four littermates each. The mice were decapitated, the skull opened down the midline and the brain removed. Newly dissected brains were rinsed in RNAse free PBS and stored at -20°C in RNAlater (Ambion). Whole brains were embedded in 5% agarose and sectioned using a vibrating microtome into 200 micrometer coronal sections using a 1:1 mixture of RNAlater and PBS. Six sections corresponding approximately to cortical layers I-III, IV, upper V, lower V, VI, and VIb (henceforth referred to as samples A-F, respectively) were dissected out under visual guidance, using transillumination on a dissecting microscope (MZFLIII, Leica) and stored separately in RNAlater at -80°C until all microdissection was complete. For RNA extraction samples from individual zones from the eight mice were combined and all tissue samples were processed at the same time. To provide a biological replicate sample, the RNA for sample B was extracted and pooled by litter, providing two samples each representing 4 mice (heretofore known as samples B1 and B2). In the same way, layers of dorsal cortex and lateral cortex were dissected from eight additional adult male mice.