Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Pool of 12 axenic larvae, L4 larvae, pupae and adult mosquitoes were surface sterilized and total RNA was extracted using Trizol. Axenic larva, L4 larva, pupa and adult mosquito libraries for controls (non-axenics) and for gnotobiotics (Flavobacterium, Enterobacteriaceae, Lysobacter and Paenibacillus treatments) were prepared from total RNA of pool of 12 individuals. Removal of ribosomal RNA, fragmentation and library preparation were performed by using the TruSeq Stranded mRNA library preparation kit (Illumina) following manufacturer's instructions. Libraries quality was checked on a DNA1000 Bioanalyzer chip (Agilent) and quantification made by QuBit DNA HS kit (ThermoFisher). single reads of 65 nucleotides in length (à verifier) were generated on a HiSeq2500 sequencing platform (Illumina).