Illumina HiSeq 2000 paired end sequencing; RNA-seq of Anopheles gambiae 24 and 48h post Trypanosoma infection
RNA-seq of Anopheles gambiae 24 and 48h post Trypanosoma infection
For each condition, a pool of 10 infected-mosquitoes has been collected at 24h and 48h post-feeding.  A. coluzzii colony Fd03 initiated in Mali was reared at 26 °C and 80 % humidity, in a 12 h light/dark cycle with access to cotton soaked in 10 % sucrose solution, in insectaries of the Unit of Genetics and Genomics of Insect Vectors of the Institut Pasteur Paris, France. Anopheles mosquitoes were fed on non-infected mice. using Illumina's Truseq RNA Sample Preparation Kit (Cat. # RS-122-2001), 1 microgram of total RNA is oligo-dT purified using oligo-dT coated magnetic beads, fragmented and then reverse transcribed into cDNA The cDNA is fragmented, blunt-ended, and ligated to indexed (barcoded) adaptors and amplified using 15 cycles of PCR. Final library size distribution is validated using capillary electrophoresis and quantified using fluorimetry (PicoGreen) and via Q-PCR.  Indexed libraries are then normalized, pooled and then size selected to 320bp +/- 5% using Caliper's XT instrument.