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Fig. 2 | Microbiome

Fig. 2

From: Targeted single-cell genomics reveals novel host adaptation strategies of the symbiotic bacteria Endozoicomonas in Acropora tenuis coral

Fig. 2

Targeted single-cell genomics with droplet microfluidics. A Microbial cells were encapsulated into 40-μm microfluidic droplets at the single-cell level (0.3 cell/droplet) with ultra-low melting temperature agarose solutions. Collected droplets were solidified, treated with lysis solution, and subjected to whole genome amplification (WGA) using a previously described single-cell amplified genome (SAG)-gel platform. After washing, whole genome amplified gel beads were resuspended in the PCR mixture. B Gel beads suspended in PCR mixture were reintroduced into microfluidic devices, and water in oil (W/O) droplets encapsulating gel beads were generated. After thermal cycling, droplets with target sequence exhibited fluorescence derived from Taqman probes. The fluorescence-positive droplets were isolated into multi-well plates and then subjected to second-round WGA and library preparation. C Microscopic images of W/O droplets after PCR. Droplets with genome amplification by WGA exhibit blue fluorescence derived from DAPI. Droplets with target gene amplification by PCR exhibit green fluorescence derived from FAM. The droplet indicated by the arrowhead contains genome amplicons of the target bacterium

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