PCR of 16s rRNA of Dugesia japonica metagenome:
Worms were briefly rinsed with sterile worm water in a petri dish to remove external microbes. Individual worms or fragments were then placed into a sterile 1.5 mL tube containing 400uL 1X PBS, using a sterile wooden dowel. Worms were then homogenized using a sterile pestle, and the homogenate serial diluted to 10-3. 80uL of each dilution was plated and spread on BHI agar plates, and plates were incubated aerobically at room temperature for five days. After incubation, bacterial colonies were grouped and tallied based on morphology. One representative colony of each morphotype was selected, DNA extracted, PCR of the 16S rRNA gene performed, the amplified product sequenced via sanger sequencing, and the identify of each morphotype confirm by classifying the sequence