title: GSM8806138: Spinal cord, distal region,10 days after injury; Mus musculus; RNA-Seq
library_construction_protocol: Single nucleus isolation and sequencing Spinal cords were rapidly extruded from iNeuroD1GFP mice (n=3) after anesthesia. Each cord was separated into lesion core and peri-injury regions. Single cell nuclei suspension of frozen tissues were prepared using the Nuclei PURE Prep kit (Sigma, NUC201). The nuclei that have passed the test are washed and resuspended to prepare a suitable cell concentration of 700~1200 nuclei/ul for 10x Genomics ChromiumTM. The system is operated on the machine. GEMs (Gel Bead in Emulsion) were constructed for single nuclei separation according to the number of nuclei to be harvested. After GEMs were normally formed, GEMs were collected for reverse transcription in a PCR machine for labeling. The GEMs were oil-treated, and the amplified cDNA was purified by magnetic beads, and then subjected to cDNA amplification and quality inspection. The 3ʹ Gene Expression Library was constructed with the quality-qualified cDNA. After fragmentation, adaptor ligation, sample index PCR, etc., the library is finally quantitatively examined.
identifiers.namespace: GEO
identifiers.id: GSM8806138_r1