Illumina HiSeq 2000 paired end sequencing; RNA-seq of Leishmania mexicana promastigotes, axenic amastigotes and intracellular amastigotes
RNA-seq of Leishmania mexicana promastigotes, axenic amastigotes and intracellular amastigotes
Bone marrow cells were harvested from C57BL/6 mice and matured in vitro in 20% L929 conditioned medium, resulting in cells expressing the murine macrophage markers F4/80 and MAC-1, whilst being negative for the granulocyte marker GR-1, as assessed by flow-cytometry. Macrophages were then cultured for 2 days in Dulbecco's Modified Eagle Medium (DMEM) before the start of the experiment. For mapping of splice acceptor sites and determination of transcript abundances, polyadenylated RNA was purified from total RNA, converted to cDNA using random hexamer primers, sheared and size selected for fragments ~200bp in length using the Illumina TruSeq RNA Sample Preparation Kit v2.
Experimental Factor: organism
Mus musculus
Experimental Factor: developmental stage
not applicable
Experimental Factor: host
not applicable (uninfected control)