GSM9138010: WT TAM,  αMARCO, rep1; Mus musculus; RNA-Seq
Total RNA was extracted and isolated following TRIzol Reagent User Guide. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. RNA integrity was assessed using the Bioanalyzer 2100 system (Agilent Technologies, CA, USA). After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. After library quality control, different libraries were pooled based on the effective concentration and targeted data amount, then subjected to Illumina sequencing by using the NovaSeq 6000 platform. As each sequencing cluster extends its complementary strand, the addition of each fluorescently labeled dNTP releases a corresponding fluorescence signal. The sequencer captures these fluorescence signals and converts them into sequencing peaks through computer software, thereby obtaining the sequence information of the target fragment.