Encephalomyocarditis virus from Loxodonta africana
Total RNA was extracted (Trizol LS)(Life Technologies, Carlsbad, CA, USA) from supernatants of infected BHK-21 cells. Samples were DNase/RNase treated prior to RNA extraction. Total RNA extract (20 -70 ng) was treated with DNase using the TURBO DNA-free kit (Ambion, Carlsbad, CA, USA) then depleted from ribosomal RNA (rRNA) using the Low Input RiboMinus Eukaryote System Kit (Ambion, Carlsbad, CA, USA). The DNA/rRNA depleted RNA solution was then converted into representative cDNA libraries, using the Ion Total RNA-Seq Kit (Life Technologies, Carlsbad, CA, USA) according to the suppliers instructions with two modifications: (i) RNA was fragmented with RNase III for 1 min at 37C and (ii) the fragmented RNA was hybridized and ligated to adaptors overnight at 16C.