title: GSM4141580: WT left M1 cortex 3; Mus musculus; RNA-Seq
library_construction_protocol: Mice were anesthetized with sodium pentobarbital (50mg/kg, intramuscularly) and either transcardially perfused with 1x PBS, followed by 4% paraformaldehyde (in 0.1 M PBS, pH 7.4) or sacrificed through cervical dislocation (for RNA extraction) and the brain was thereafter dissected out. Left and right primary motor cortex was biopsied either by needle puncture (for RNA-sequencing) or using small dissection forceps (approx. 1.0 x 1.0 mm pieces for other biochemical analyses) and put individually in tubes containing RNAlater solution (Invitrogen). The tubes were stored at -80°C before RNA extraction. Total RNA was isolated using the RNeasy Plus Micro kit and the RNeasy kit respectively (both from Qiagen, Hilden, Germany) according to manufacturer's recommendations. 10 ng of RNA from left and right primary motor cortex biopsies from 6 WT and 6 BERKO mice were processed using the Single-cell Tagged Reverse Transcription (STRT) RNA sequencing protocol. Briefly, RNA samples were placed in a 48-well plate in which a universal primer, template-switching helper oligos, and a well-specific 6 bp barcode sequence (for sample identification) were added to each well. Reverse transcription reagents and ERCC RNA Spike-In Mix I (both from Thermo Fischer Scientific) were added to generate first-strand cDNA. The synthesized cDNAs from the samples were then pooled into one library and amplified by single-primer PCR with the universal primer sequence. The resulting amplified library was then sequenced in 3 lanes using Illumina HiSeq 2000 instrument.
attributes.tag: GEO Accession
attributes.value: GSM4141580