title: GSM6203349: BM from control mouse 1 [5-1]; Mus musculus; RNA-Seq
library_construction_protocol: BM cell suspensions were treated 529 with Diisopropylfluorophosphate (1mM) prior to cell sorting. Cell suspensions were stained with monoclonal antibodies to mouse CD45.1, CD45.2, CD11b, F4/80 and single cell macrophages (CD45.1+ CD11b+ F4/80+) were sorted into 96 well-plates containing 0.2% Triton X-100 and 0.1ul of RNAase plus inhibitor (Promega N2615)/each well using BD FACS ARIA Z6001 (BD Biosciences). In brief, oligo-dT30 primers and dNTP mix were mixed with isolated RNA and incubated with samples at 72°C for 3 min. Reverse transcription and pre-amplification steps were performed. cDNA was then purified, and library quality assessed using Tapestation D5000. Samples were subsequently tagmented and amplified according to the Nextera XT DNA Library Preparation protocol. Indexed samples were then pooled and cleaned. Libraries were then sequenced at 2x75bp paired end reads, with a total of 100M reads.