GSM7057446: WT control; Mus musculus; RNA-Seq
whole lungs were minced and digested with 500 μg/mL Liberase TB (Sigma, # 5401127001), 150 μg/mL DNase I (Sigma #1010459001), and 5 mM MgCl2 in PBS (pH 7.4) for 20 min at 37 oC. Then, single cell suspension was filtered through 70 μm nylon mesh cell strainer and treated with RBC Lysis Buffer (Invitrogen, # 00430054) according to manufacturer's instructions. Ten million lung cells in suspension were mixed with 5 μL Component A, 5 μL Component B, and 5 μL magnetic RapidSpheresTM provided in the EasySep kit, and incubated for 5 min in ice. CD45+ cells trapped in magnetic beads were isolated using a magnetic platform, and released diluted in EasySepTM Buffer (StemCell #20144). Sample quality, library complexity, and alignment statistics were checked using an established pipeline at the NUSeq Core Facility at Northwestern University