V4 region of 16S rRNA metagenome of ZymoBIOMICS Microbial Community Standard - Illumina run rep a
Microbiome samples from each Symbiodiniaceae culture, as well as negative controls, were collected at about the same time of day. Only bacteria that were physically attached to the Symbiodiniaceae cell surface were sampled. DNA extraction was performed using the DNeasy PowerSoil Pro kit (Qiagen) according to the manufacturers protocol. Three replicates of a mock community, the ZymoBIOMICSMicrobial Community Standard (Zymo Research), were included as positive controls. The V4 region of the 16S rRNA gene was amplified in 20 l PCR reactions. A sample of autoclaved nuclease-free water was included as a negative PCR control. PCR products were submitted to the Rush University Medical Center Genomics and Microbiome Core Facility for library preparation, paired-end sequencing on an Illumina MiSeq i100+, and sequence processing.