GSM7385465: WT_HFD bio rep 2; Mus musculus; RNA-Seq
Mouse lung endothelial cells were isolated by mincing and digesting lung tissue in 1mg/ml collagenase dispase, filtering the suspension through 70um and 30um strainers followed by selection with anti CD31 antibody-conjugated microbeads and MACS separator  (Miltenyi). RNA was isolated using a Promega Reliaprep RNA extraction kit. Libraries were prepared from 3 biological replicates per group. mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis. cDNA underwent 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.