16s profiling of Ochlerotatus taeniorhynchus: whole body
Samples were amplified via PCR with the 515F/806R primers for the V4 region of the 16S rRNA gene, and using the HotStarTaq Plus Master Mix Kit (Qiagen). Forward primers were barcoded in order to permit sample pooling. PCR conditions were as follows: 3 mins at 94C, followed by 30 cycles of 30 sec at 94C, then 40 sec at 53C, then 60 sec at 72C, and finally 5 mins at 72C. PCR products for all samples were then run on 2% agarose gels in order to gauge band size and intensity. Amplified DNA for each sample was purified using Ampure XP beads. Samples were pooled in equal proportions, according to the concentration of DNA and the molecular weight, and this mixture was used to prepare Illumina DNA libraries.