16S rRNA microbiome sequencing of CeMbio bacteria from gut and skin of C. elegans
16S rRNA sequences acquired from pooled C. elegans raised on CeMbio bacterial strains exposed to surface sterilization (bleaching) protocols or serial washes to explore the bacteria on the C. elegans cuticle. Specifically, C. elegans were raised on CeMbio bacterial lawns. Adult worms were washed with M9 and picked off using sterile pipettes. For serially washed ("nobleach") animals, groups of 30 animals were washed 5 times with M9. Bleached ("bleach") animals were collected similarly and washed washed with bleaching solution (M9 with 25mM tetramisole, 2%bleach,1M KOH) and a total of 4 times with M9. For each condition, 100uL of M9 and 30 C. elegans were frozen at -80C. "Lawn" represents scraped bacterial slurry in 1mL M9 without any animals present, centrifuged and frozen in 100uL of M9. DNA was extracted using NucleoSpin Tissue kit protocol for Animal Tissue with 1 hr of incubation for the first lysis step. DNA was stored at -20C.