RNA-Seq of Homo sapiens: flow-sorted T cells from bronchoalveolar lavage fluid
     Following sorting of T cell subsets, cells were pelleted by centrifugation and      lysed in 350 μl of RLT Plus lysis buffer (Qiagen) supplemented with 1% 2-mercaptoethanol      and immediately stored at −80 °C. A Qiagen AllPrep DNA/RNA Micro kit was used      for simultaneous isolation of RNA and DNA, and RNA quality and quantity was assessed      using a 4200 TapeStation System (Agilent Technologies). RNA-sequencing libraries were      prepared from 300 pg of total RNA using a SMARTer Stranded Total RNA-seq Kit v2      Pico Input Mammalian (Takara Bio). Libraries were pooled and sequenced on a      NextSeq 500 instrument (Illumina; 75 cycles, single end) to an average      sequencing depth of 20.83 million reads.     
NIAID-phs002300
S360795734_S9