title: 16S rRNA population analysis of Ariolimax fecal microbiome
design: Slug fecal samples were collected and stored in TE buffer at -80 C until all were ready for sequencing. DNA from each fecal homogenate sample was extracted with a DNeasy PowerSoil Kit (Qiagen). Each PowerBead sample was disrupted with a Mini-beadbeater (Biospec Products, Bartlesville, OK), with the power set to 42 Hz twice for 30 seconds. Tubes were chilled on ice 1 minute after each beadbeating. Subsequent extraction procedures followed the manufacturers instructions (Qiagen), and all DNA samples were eluted in a final 100 l of Solution C6 and stored at -80C. All isolated DNA samples were amplified and analyzed by the Research and Testing Laboratory (RTL; Lubbock, TX) using Illumina MiSeq analysis. Amplicons of the V1-V2 hypervariable regions of the 16S rRNA gene were generated using the primers 28F (5-GAGTTTGATCNTGGCTCAG-3) and 388R (5-TGCTGCCTCCCGTAGGAGT-3)