GSM2720222: s91931_M5CRe-; Mus musculus; RNA-Seq
CD45+ Ly6G– CD11b+ F4/80+ cells were sorted into cold PBS containing 5% BSA and were sedimented by centrifugation. An aliquot of the sorted cells was subjected to reanalysis to determine the degree of sorting purity. Cells were resuspended in 350 μl of RLT+b-ME buffer (RNeasy Micro Kit, Qiagen), stored at -80°C and used for Illumina-based sequencing. RNA quantity and quality was determined by measurement on the Qubit device as well as using the Agilent BioAnalyzer 2100. Only the samples with the RNA integrity number above 8 were sequenced. SMARTer Ultra Low Input RNA for Illumina Sequencing Kit - HV from Clontech was used to reverse transcribe the RNA and amplify full-length cDNA according to the user manual. It was followed by a library prep using NEBNext Ultra DNA Library Prep Kit for Illumina (NEB).