GSM9329367: CD11B MUT vehicle rep2; Mus musculus; RNA-Seq
Myeloid CD11b⁺ cells were isolated by FACS 7 days after injury. For RNA isolation from FACS-isolated CD11B-positive cells, the RNeasy Plus Micro Kit (Qiagen, 74034) was employed according to the manufacturer's instructions. P3422 : RNA-Seq libraries were generated from 50 ng of total RNA using Illumina Stranded mRNA Prep, Ligation kit and IDT for Illumina RNA UD Indexes Ligation (Illumina, San Diego, USA) according to manufacturer's instructions. Following polyA selection, mRNAs were fragmented at 94oC for 2 minutes. DNA library were amplified using 14 cycles of PCR. Surplus PCR primers were further removed by two successive purifications using SPRIselect beads (Beckman-Coulter, Villepinte, France).  The final libraries were checked for quality and quantified using Bioanalyzer 2100 system (Agilent technologies, Les Ulis, France).