title: V3-V4 regions of the 16S rRNA gene of Bacteria: Anopheles vagus abdomens
design: DNA from pooled larvae and adult abdomens was extracted using the NucleoSpin tissue kit (Macherey-Nagel GmbH & Co. KG, Duren, Germany) according to the manufacturer's instructions. DNA concentration and purity were monitored using a NanoDrop Spectrophotometer and electrophoresis through 1% agarose gels. The next-generation sequencing analysis was based on the 16S rRNA gene of bacteria. The V3-V4 regions of the gene were amplified using universal region-specific primers 341F (5-CCT AYG GGR BGC ASC AG-3) and 806R (5-GGA CTA CNN GGG TAT CTA AT-3) tagged with sample-identifying barcodes. PCR products of the proper size were selected by 2% agarose gel electrophoresis. The DNA fragments were end-repaired and A-tailed then further ligated with Illumina adapters. The libraries were generated and sequenced on a paired-end Illumina platform.