Additional file 6 of Bacterial communities in carnivorous pitcher plants colonize and persist in inquiline mosquitoes

Additional file 6: Fig. S 4. PCR analysis of W. smithii larvae under axenic, gnotobiotic, or conventional conditions. Axenic first instars from surface-sterilized eggs were hatched in closed containers containing sterile water and sterilized diet. Gnotobiotic larvae recolonized by their native microbiota were produced by feeding axenic larvae sterilized diet plus material from a glycerol stock containing the mixed community of bacteria present under conventional rearing conditions. Conventional first instars were hatched in open containers containing distilled water and standard diet. For each treatment, DNA was isolated from a pooled sample of at least 10 larvae after surface sterilization as described herein (see ���Methods���). DNA samples were then used as template with universal bacterial 16S rRNA gene or fungal ITS primers. The agarose gel shows ethidium bromide-stained PCR products. Lane 1, molecular mass markers labeled in base pairs (bp); Lanes 2���4, universal 16S rRNA gene primers plus DNA from axenic larvae; Lanes 5���7, universal ITS primers plus DNA from axenic larvae; Lanes 8���10, universal 16S rRNA gene primers plus DNA from gnotobiotic larvae; Lanes 11���13, universal 16S rRNA gene primers plus DNA from conventional larvae.