title: GSM4478867: EV_Brain_5; Mus musculus; RNA-Seq
library_construction_protocol: Substantia Nigra and striatum were microdissected, incubated for 30 min at 37°C and 650 rpm in 1mL HBSS with 5 mg Collagenase NB4 and 0.1 mg DNaseI followed by the generation of cell suspensions in HBSS with a 70 µm cell strainer. Cells were centrifuged at 240 g and 18°C for 5 min. The pellet was resuspended in 30% Percoll/PBS and immune cells were isolated after centrifugation at 650G and 18°C for 30 min with a 70% Percoll/PBS gradient. RNA isolation was performed with the RNeasy micro kit (Qiagen cat # 74004) according to manufacturer's instructions. Libraries were prepared from 0.5 ng of total RNA using the SMARTer Stranded Total RNA-Seq Kit v2 (Takara Bio cat#634411) with 4 min RNA fragmentation at 94°C, 5 cycles of PCR for cDNA library pre-amplification and 15 cycles of PCR amplification after rDNA depletion.
attributes.tag: GEO Accession
attributes.value: GSM4478867