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: Axenic amastigotes were generated by dilution of stationary phase Leishmania mexicana promastigotes (WHO strain MNYC/BZ/62/M379) into Schneider’s Drosophila medium at pH 5.5 to a density of 3x10E6 cells/ml and incubation at 34°C for 24h prior to RNA extraction. For determination of polyadenylation addition sites, polyadenylated RNA purified from total RNA was converted to cDNA using a 5’-T15VN-3’ oligonucleotide (V=A, G or C; N=T, A, G or C) primer to enrich for 3’ ends of transcripts. The cDNA was 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: axenic amastigote
attributes.tag: Experimental Factor: host
attributes.value: not applicable (outside host)