fecal water control sample from cage 233 - 1
DNA sample were extracted either with the QIAamp DNA Stool Kit (Qiagen) or a DNA extraction protocol developed for C. elegans  (Zhang F, et al. (2020)).   Supernatants from lysates were transferred to a clean 96-well PCR plate and used as DNA templates. We amplified the V4 region of the 16S rRNA gene using the 515f (GTGYCAGCMGCCGCGGTAA) and 806r (GGACTACNVGGGTWTCTAAT) PCR primers with barcodes added to the 806r primer. Amplicons for each library were normalized based on the PCR product quantified by the image processing package in Fiji (50, 51), then pooled into a single tube. Amplicon products were quantified (Qubit) and then sequenced by the Center for Metagenomics and Microbiome Research, Houston, Texas, USA on an Illumina MiSeq System.