GSM4629807: Infection_3_RNAseq_salivary_glands_paired; Anopheles gambiae; RNA-Seq
We performed dissection of midguts on adults at 7 days post-infection and of salivary glands at 14 days post-blood meal. Total RNA was extracted from fresh mosquito tissues (~30 midguts and ~60 salivary glands). Immediately after dissection, tissues were stored in TRIzol (Invitrogen) and frozen at -80ºC until subsequent processing. We used the Ovation® Universal RNA-seq System (Nugen Technologies) for strand specific RNA-seq library construction following the manufacturer's instructions. Custom primers specific to mosquito ribosomal sequences were designed to reduce the percentage of ribosomal reads in the sample and the ribo-depletion step was incorporated into the standard workflow. RNA concentration was quantified using a Qubit® 2.0 Fluorometer, and RNA integrity was determined with an Agilent 2100 Bioanalyzer. Illumina libraries were prepared and sequenced at the Cabimer, using an Illumina NextSeq500 sequencer, 75 bp single-end reads.
GEO Accession
GSM4629807