GSM7661272: BCG+ Celecoxib- Rv+ [R12649]; Mus musculus; RNA-Seq
Total RNA was extracted from flow-sorted lung neutrophils from Mtb-infected WT mice, treated or not with BCG and/or Celecoxib, following a double extraction protocol: RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction (TRIzol, Thermo Fisher Scientific, Waltham, MA, USA) followed by a Qiagen RNeasy Micro clean-up procedure (Qiagen, Hilden, Germany). cDNA libraries were prepared using 0.5 ng of total RNA using the Smart-seq2 protocol with the following modifications: 1. Addition of 20 µM TSO; 2. Use of 200 pg cDNA with 1/5 reaction of Illumina Nextera XT kit (Illumina, San Diego, CA, USA). The length distribution of the cDNA libraries was monitored using a DNA High Sensitivity Reagent Kit on the Perkin Elmer Labchip (Perkin Elmer, Waltham, MA, USA). All samples were subjected to an indexed paired-end sequencing run of 2x151 cycles on an Illumina HiSeq X system (28 samples/lane).