title: GSM9220470: MG_C_141_JJ; Rattus norvegicus; RNA-Seq
library_construction_protocol: To study differences in microglial gene expression, bulk RNA sequencing was performed. RNA from microglial cells was isolated using the RNeasy Micro Kit (Qiagen, 74104), according to the manufacturer's guidelines. mRNA was enriched from total RNA with the Dynabeads® mRNA Purification Kit (Invitrogen, 61006). A strand-specific transcriptome library was constructed using the MGIEasy RNA Directional Library Prep Set (16 RXN) (MGI, 1000006385). Briefly, after cDNA synthesis, adaptor ligation was performed to ligate adaptors with the cDNAs. Subsequently, a PCR reaction was conducted to amplify the cDNAs, after which the library quality was checked using the Agilent 2100 Bioanalyzer (Agilent, G2939AA). Finally, single-stranded cyclized products were produced from single-stranded PCR products. To perform sequencing, single-stranded circle DNA molecules were replicated via rolling cycle amplification and a DNA nanoball (DNB) was generated. Sequencing was performed through combinatorial Probe-Anchor Synthesis (cPAS) on the DNBSEQ-G400 (MGI, G400).