RNA-Seq of Homo sapiens: flow-sorted macrophages from bronchoalveolar lavage fluid: Sample 340228397
Immediately after sorting, cells were pelleted by centrifugation and lysed in 350 μl RLT Plus lysis buffer (Qiagen) supplemented with 2-mercaptoethanol. Lysates were stored at −80 °C until RNA isolation using the AllPrep DNA/RNA Micro kit according to the manufacturer's protocol (Qiagen). RNA quality and quantity were assessed using TapeStation 4200 High Sensitivity RNA tapes (Agilent), and RNA-seq libraries were prepared from 250 pg of total RNA using SMARTer Stranded Total RNA-seq Kit v2 (Takara Bio). Libraries were pooled using dual indexing and sequenced on a NextSeq 500 instrument (Illumina), 75 cycles, single-end, to an average sequencing depth of 19.55 million reads.