GSM5456868: Spinal motor neurons, Wild-type, N268; Mus musculus; RNA-Seq
Lumbar spinal cords were dissected and immediately snap-frozen in 2-methylbutane cooled in liquid nitrogen. Spinal cords were embedded in OCT, cryosectioned at −20°C (12 μm coronal sections) onto PEN membrane glass slides (Zeiss) and subsequently stored at −80°C until further processing. The sections were then used for laser capture microdissection of spinal motor neurons as described in Nichterwitz 2016. Briefly, the sections were thawed 30 seconds, subjected to quick histological staining (Histogene, Arcturus/LifeTechnologies) and cells were captured using the Leica LMD7000 system at 40x magnification. Only cells with a visible nucleus with nucleolus and with an area of more than 200 μm2 were captured. Cells were directly lysed with 5 μl lysis buffer (0.2% Triton X-100, with 2 U/μl recombinant RNase inhibitor, Clontech) Libraries for RNA-seq were prepared using SMART-seq2 combined with Tn5 tagementation
GEO
GSM5456868