title: GSM4143989: TG1; Mus musculus; RNA-Seq
library_construction_protocol: PBS perfused kidneys were minced and digested with Liberase TL (0.25 mg/ml) and RNAse free DNAse 1 (100u/ml) in DMEM with rotation at 225 rpm at 37°C for 30 minutes. Tissue digests were filtered through a 40 um nylon mesh (BD Bioscience) and washed with PBS. Dead cells were removed with an immuno-magnetic Dead Cell Removal Kit (Miltenyi, Auburn, CA). 1 X 107 live cells were incubated with anti-mouse F4/80 magnetic beads (Miltenyi) for positive selection of MPC cells. Pelleted cells were lysed in RLT buffer followed by RNA isolation using an RNeasy micro kit with gDNA eliminator columns (Qiagen). The library preparation was done using a NEBNext Ultra II Directional RNA Library kit (New England Biolabs).  75 ng of total RNA was mRNA enriched using oligodT beads. The mRNA-stranded libraries were pre-amplified with PCR incorporating dual index primers and purified with magnetic beads.
attributes.tag: GEO Accession
attributes.value: GSM4143989