title: GSM6436853: bone marrow-derived macrophages cultured in 50% LL-2 conditional medium 24h 1; Mus musculus; RNA-Seq
library_construction_protocol: BMDMs were harvested and lysed in TRIzol (Invitrogen). The extraction and sequencing of RNA was performed by Novogene Bioinformatics Institute, Beijing, China. After the RNA samples passed the test, eukaryotic mRNA was enriched with magnetic beads with Oligo(dT) (in case of prokaryotes, mRNA was enriched by removing rRNA with the kit) . The mRNA was then broken into short fragments by adding fragmentation buffer, and one-stranded cDNA was synthesized using six-base random primers (random hexamers) with mRNA as the template, followed by the addition of buffer, dNTPs and DNA polymerase I and RNase H to synthesize two-stranded cDNA, followed by the use of The purified double-stranded cDNA was firstly end-repaired, A-tailed and sequenced with AMPure XP beads, and then fragment size selection was performed with AMPure XP beads. Finally, PCR amplification was performed, and the PCR products were purified with AMPure XP beads to obtain the final library. After library construction, initial quantification was performed using Qubit 2.0 to dilute the library, followed by detection of the insert size of the library using Agilent 2100, and after the insert fragment met the expectation, the effective concentration of the library was accurately quantified using Q-PCR method to ensure the quality of the library.