16S rRNA sequencing of Stichopus monotuberculatus: regenerating gut samples
Total DNA was extracted from intestinal tissue samples using Qiagen kits according to the manufacturer's protocols. The V3-V4 region of the 16S rRNA gene was PCR-amplified using primers 341F and 806R. Sequencing libraries were prepared with the NEBNext Ultra II DNA library prep kit and sequenced on the Illumina Novaseq6000 platform, generating paired-end reads. Raw reads were processed using FLASH for merging and QIIME2 for clustering into OTUs, with taxonomy assigned via the SILVA 132 database.