GSM9743656: primary microglia from CD31-KD mice after 24-hour treatment with Aβ oligomers-3; Mus musculus; RNA-Seq
Total RNA was extracted from primary microglial cells treated with Aβ oligomers for 24 hours, including both control and CD31 knockdown groups. mRNA enrichment was carried out using magnetic beads with Oligo(dT) to selectively isolate polyA-tailed mRNA. The enriched mRNA was fragmented using a fragmentation buffer, followed by reverse transcription with random N6 primers to synthesize the first-strand cDNA. The second cDNA strand was synthesized to generate double-stranded DNA, which was end-repaired, 5′ phosphorylated, and an 'A' was added to the 3′ end. The DNA fragments were ligated to a bubble-like adapter with a protruding 'T' at the 3′ end. PCR amplification was performed with specific primers, and the PCR products were denatured into single strands. These strands were circularized using a bridging primer to form a single-stranded circular DNA library. Library quality was assessed, and sequencing was performed after passing quality control.