Healthy sea urchin coelomic fluid
The gDNA samples were processed, sequenced, and analyzed using the ZymoBIOMICS Targeted Sequencing Service at ZymoResearch (Irvine CA). The targeted sequencing of the bacterial 16S rRNA gene was performed using the Quick-16S NGS Library Prep Kit (ZymoResearch) using custom-designed primers to amplify the V3-V4 region of the 16S rRNA gene. The library preparation was completed by real-time PCR and quantification of the PCR products using qPCR fluorescence readings. Library fragment sizes were selected and optimized with the Select-a-Size DNA Clean & Concentrator (ZymoResearch) and quantified with Tapestation (Agilent Technologies) and Qubit (ThermoFisher Scientific). The positive control sample used for library preparation was the ZymoBIOMICS Microbial Community DNA Standard (ZymoResearch) The completed library was sequenced using a V3 reagent kit with 600 cycles on Illumina MiSeq, which was calibrated by a 10% spike-in of PhiX DNA.