title: Illumina HiSeq 2000 sequencing; gambiae_RNAseq
design: gambiae_RNAseq
library_construction_protocol: The Anopheles gambiae Ngousso strain was originally initiated with mosquitoes collected in Cameroon in January 2006, and belongs to the M molecular and Forest chromosomal forms (Harris C, Lambrechts L, Rousset F, Abate L, Nsango SE, Fontenille D, Morlais I, Cohuet A. (2010). Polymorphisms in Anopheles gambiae immune genes associated with natural resistance to Plasmodium falciparum. PloS Pathog 6, e1001112.). Mosquito were reared under standard conditions as previously described (Mitri C, Jacques JC, Thiery I, Riehle MM, Xu J, Bischoff E, Morlais I, Nsango SE, Vernick KD, Bourgouin C. (2009). Fine pathogen discrimination within the APL1 gene family protects Anopheles gambiae against human and rodent malaria species. PloS Pathog 5, e1000576. Mosquitoes were infected on an infectious bloodmeal containing physiologically natural viral titers O'Nyong-Nyong strain SG650 12 mosquitoes per pool, in each of two replicates, were collected 3 days after bloodmeal infection with ONN and homogenized in TriReagent (Sigma). Small RNAs fraction (<200 nt) were extracted using Nucleospin miRNA (Machery Nagel) using the Trizol protocol and submitted to Bioanalyser for quality assessment. Briefly, the 10- to 30-nt RNA fragments were purified from small RNA fraction on a 15% urea PAGE (Biorad). Small RNA library were prepared using TruSeq Small RNA Library reagents (Illumina).
attributes.tag: Experimental Factor: infect
attributes.value: o'nyong'nyong virus