GSM7872696: AAEFMG, replicate1; Aedes aegypti; RNA-Seq
Female adult mosquitoes at 7 days after eclosion were anesthetized on ice. Subsequently, midguts were dissected from a group of 40 mosquitoes, then washed and temporarily stored in a microtube filled with Schneider's Drosophila Medium kept on ice. After the dissection, the medium was discarded, and 400 ul of dissociation buffer was added into the microtube. The mixture was incubated on a rotator for 20-30 min at room temperature. Then, 400 ul of Schneider's Drosophila Medium was added to the microtube and mixed well. A total of 800 ul of the cell suspension was filtered with a 40 um Flowmi Cell Strainer to remove larger cell debris. To remove impurities and smaller cell debris, the filter was centrifuged over a density of approx. of 1.12 g/ml made with OptiPrep at 400× g RCF, 4 C for 20 min. Viable cell band was collected using a 200 ul pipette tip, mixed well in a new microtube and centrifuged at 400× g for 20 min again. The cell precipitation was washed with 1× PBS for several times and then resuspended with 200 ul of Schneider's Drosophila Medium. Library construction was performed with Single Cell 3' v3 Chemistry according to the manufacturer's instructions.