PacBio-based sequencing of the Anopheles gambiae 4 h-old male and female pupae transcriptomes

The study aims at exploration of transcriptome of the Anopheles gambiae 4 h-old pupae. To better understand exon connectivity in longer, multi-exon transcripts, PacBio platform was used for sequencing. Total RNA was extracted from batches of males or females, followed by isolation of poly-A (+) fraction using NEB Magnetic mRNA Isolation kit and cDNA synthesis. Sex-specific cDNA libraries were subjected to double size selection using AMPure XP beads (0.4x and 1.0x), and, for each sex, the “fractions” were pooled in equimolar ratios for a single SMRTbell library construction. The libraries were sequenced on a Sequel instrument.