title: Illumina HiSeq 2000 paired end sequencing; RNA-seq of Leishmania mexicana promastigotes, axenic amastigotes and intracellular amastigotes
design: RNA-seq of Leishmania mexicana promastigotes, axenic amastigotes and intracellular amastigotes
library_construction_protocol: Promastigote-form L. mexicana (WHO strain MNYC/BZ/62/M379) were grown at 28°C in M199 medium (Life Technologies) supplemented with 2.2 g/L NaHCO3, 0.005% haemin, 40 mM 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES) pH 7.4 and 10% FCS (Life Technologies 10500-064). Cells were maintained in exponential growth phase prior to RNA extraction. 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.
attributes.tag: Experimental Factor: organism
attributes.value: Leishmania mexicana
attributes.tag: Experimental Factor: developmental stage
attributes.value: promastigote
attributes.tag: Experimental Factor: host
attributes.value: not applicable (outside host)