GSM4221615: Aedes_Diro.Lvp.d1.1; Dirofilaria immitis; Aedes aegypti; RNA-Seq
Malpighian tubules were dissected into PBS by pulling at the last abdominal segment with forceps until the hindgut emerges. Next, the abdomen is opened where it joins the thorax and the midgut was separated from the thorax. The hindgut, midgut, and Malpighian tubules are removed from the posterior abdomen by pulling again on the previously separated posterior segment with forceps. The hindgut and midgut were separated, leaving a set of tubules. Tubules from five mosquitoes were transferred together to a tube containing Trizol (Life Technologies) and stored at -80 ºC until processing according to the manufacturer's instructions. Purified total RNA was resuspended in water and treated with Turbo DNA-free (Invitrogen) to remove potentially contaminating genomic DNA. The RNA concentration and quality were assayed using a NanoDrop (ThermoFisher), 4200 TapeStation (Agilent) and Qubit 3 Fluorometer (ThermoFisher). Overall, 23 libraries were generated from the 24 conditions, as one replicate of Ae. aegyptiS control did not yield a sufficient quantity of RNA for library production RNA-seq libraries were prepared using the TruSeq Stranded mRNA Library Prep Kit according to the 9 manufacturer's instructions. Samples were run on Illumina NextSeq 500 using a two High Output v2 kit flow cells to generate 75-base pair, single-end reads.
GEO
GSM4221615
GEO Accession
GSM4221615