miseq barcoded sequencing for 16s rrna gene in bacteria and its region in fungi  (Illumina Inc)

Journal: Current Biology

Article Title: Stable high-density and maternally inherited Wolbachia infections in Anopheles moucheti and Anopheles demeilloni mosquitoes

doi: 10.1016/j.cub.2021.03.056

Figure Lengend Snippet: FISH Wolbachia visualization in the ovaries Wolbachia was primarily located in the ovarian follicles (A–H). Colored boxes indicate area of magnification for subsequent images. Within the same ovary, some ovarian follicles are sparsely infected with Wolbachia (E and magnification in G), while others have a heavy infection (C, D, and H; E and F). Asterisks indicate infection in the secondary follicles. Wolbachia was imaged with an Alexa 590-labeled probe targeting the Wolbachia 16S rRNA gene (red), and DNA was stained with DAPI (blue). No probe control images (I–L) show no fluorescent signal (images in I and J and in K and L are for two separate individuals). FISH analysis revealed that 9/16 individuals were Wolbachia infected. See also Figure S2 .

Article Snippet: The microbiomes of selected individual mosquitoes were analyzed using barcoded high-throughput amplicon sequencing of the bacterial 16S rRNA gene (with library preparation and Illumina sequencing carried out commercially by Source Bioscience, Cambridge, UK).

Techniques: Infection, Labeling, Staining, Control

Journal: Current Biology

Article Title: Stable high-density and maternally inherited Wolbachia infections in Anopheles moucheti and Anopheles demeilloni mosquitoes

doi: 10.1016/j.cub.2021.03.056

Figure Lengend Snippet: Wolbachia strain densities and relative abundance in the mosquito microbiome (A) Normalized Wolbachia strain densities measured using qPCR of the conserved Wolbachia 16S rRNA gene. A synthetic oligonucleotide standard was used to calculate Wolbachia 16S rRNA gene copies per nanogram of total DNA using a 10-fold serial dilution standard curve. p values from t tests are shown to indicate significant differences. (B) Relative Wolbachia abundance in the mosquito microbiome. Taxonomic abundance of bacterial ASVs within the 16S rRNA microbiomes of An. demeilloni and An. moucheti using QIIME 2 was used to determine Wolbachia percent abundance of total 16S rRNA bacterial load, indicated through box-and-whisker plots (GraphPad Prism 9). See also Figure S3 .

Article Snippet: The microbiomes of selected individual mosquitoes were analyzed using barcoded high-throughput amplicon sequencing of the bacterial 16S rRNA gene (with library preparation and Illumina sequencing carried out commercially by Source Bioscience, Cambridge, UK).

Techniques: Serial Dilution, Whisker Assay

Journal: Current Biology

Article Title: Stable high-density and maternally inherited Wolbachia infections in Anopheles moucheti and Anopheles demeilloni mosquitoes

doi: 10.1016/j.cub.2021.03.056

Figure Lengend Snippet:

Article Snippet: The microbiomes of selected individual mosquitoes were analyzed using barcoded high-throughput amplicon sequencing of the bacterial 16S rRNA gene (with library preparation and Illumina sequencing carried out commercially by Source Bioscience, Cambridge, UK).

Techniques: SYBR Green Assay, Sequencing, Software