title: GSM6281279: RK010, scRNAseq; Rattus norvegicus; RNA-Seq
library_construction_protocol: All tissues were homogenized using a Minute Single Nucleus Isolation Kit for Neuronal Tissues/Cells (Invent Biotech. BN-020) single-nucleus suspensions were mixed with Single Cell Master Mix containing template switch oligonucleotide and loaded together with 3′ gel beads and partitioning oil into a Chip G according to the manufacturer's instructions (10x Genomics). RNAs within single nuclei were uniquely barcoded and reverse transcribed within droplets. The RT reaction was performed on a Veriti PRO Thermal Cycler (Applied Biosystems) at 53°C for 45 minutes. Then, cDNAs were amplified using cDNA primer mix from the kit via PCR for 13 cycles, and then the manufacturer's instructions were followed. Each library was barcoded with different indexes from a Dual Index Kit TT Set A plate (10x Genomics). The size profiles of the libraries were examined by a Bioanalyzer (Agilent), and the amount of RNA was quantified by KAPA Library Quantification Kits (Kapa Biosystems). All libraries were sequenced on the HiSeq X platform (Illumina) as 150 bp paired-end reads.