GSM5600340: 8A; Mus musculus; RNA-Seq
Brains were divided into 1mm-thick slices using brain matrix and razor blades that were kept chilled with dry ice. Peri-infarct regions of primary motor and somatosensory cortex, homotopic regions in the contralesional cortex or corresponding uninjured cortex in naïve animals were dissected from the resulting slices using 1 mm ID tissue micropuncher (Reusable Rapid Punch kit, WPI, Hitchin, UK) in a setup that was continuously chilled with dry ice and kept at -80°C until use. Tissue micropunches were lysed with Qiazol reagent (Qiagen, Hilden, Germany) and homogenized by passing the lysate through microsyringe fitted with 23G needle 10 times followed by repeating the above procedure using a 27G needle. Total RNA was extracted with RNAeasy Mini Lipid kit (Qiagen), including on-column DNAase I digestion step, according to manufacturer's recommendations. The RNA-Seq libraries were prepared from 100 ng of Total RNA using the QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina (Lexogen).