RNA-seq Profiling of M. marinum Infection

Two separate M. marinum infection experiments were prepared for RNA-seq. Total RNA was isolated by the Ambion PureLink™ RNA Mini kit in combination with TRIzol (Invitrogen), followed by on column DNase treatment according to the manufacturer’s instructions (Ambion, Invitrogen). RNA yield and quality were measured by Nanodrop (Thermo Scientific) and by agarose gel electrophoresis, respectively. Preparation of L. pneumophila infected samples were described previously [23 (link)]. Total RNA samples, biological duplicates for each time point, were sent to SciLifeLab Stockholm for library preparation and high-throughput sequencing. mRNA was isolated from the total RNA pool using poly(A) extraction with oligo dT prior to sequencing with Illumina HiSeq 2000/2500 system.