title: GSM9566206: Primary microglia, LPS, 4 h, replicate 1; Rattus norvegicus; RNA-Seq
library_construction_protocol: Total RNA was extracted from primary microglia, and its integrity was determined using agarose gel electrophoresis. The concentration of RNA was accurately determined with a NanoDrop spectrophotometer. Magnetic beads coated with oligo-dT were employed to isolate mRNA from total RNA. The captured mRNA was fragmented, and the first and second strands of cDNA were then synthesized using reverse transcriptase. The cDNA fragments were processed using an end-repair reaction after the addition of a single “A” base, followed by adapter ligation. These ligation products were purified and then amplified by PCR to generate the final cDNA library, which was further purified with magnetic beads. After library preparation was complete, the concentration was detected with a Qubit instrument, and the length of the library fragments was detected with an Agilent fragment analyzer to ensure quality.