title: Stranded mRNA-Seq of Bos Taurus calf, Animal 847, challenged, time 4 weeks.
design: Determination of transcriptional signatures in bovine whole blood that result from a Mycobacterium bovis infection. Two to three month old Holstein steers were acclimated for two months. Animals were split into two groups, control and challenge.  Whole blood was drawn (time 0) from each animal immediately before the challenge group was exposed to aerosolized Mycobacterium bovis strain 10-7428.  Additional blood draws were made at 4 and 10 weeks post challenge, time 4 and 10, respectively.  The LeukoLOCK Fractionation & Stabilization Kit (Ambion, Life Technologies) was used to isolate total leukocytes from whole blood. Filters were rinsed with phosphate buffered saline followed by RNAlater (Ambion), and stored at -80C until the time of RNA processing.  TRI Reagent (Life Technologies) was used to isolate RNA from the leukocyte containing filters according to the alternative extraction protocol for cells captured on LeukoLOCK filters.  The eluted RNA was treated with DNase from the DNA-free Kit (Ambion, Life Technologies).  RNA samples were stored at -80C.  RNA samples were checked for purity and quantity and then prepared into libraries using the Universal Plus mRNA-Seq Kit (NuGEN Technologies).  The stranded mRNA libraries were sequenced on an Illumina HiSeq3000 using the 2 x 100 bp paired-end methodology.