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MB Sample ID: SA029930
Local Sample ID: | 1_CL5 |
Subject ID: | SU000592 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL/6 |
Species Group: | Mammal |
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Treatment:
Treatment ID: | TR000606 |
Treatment Summary: | Animals Specific-pathogen-free (SPF) C57BL/6 mice were originally purchased from Jackson Laboratories. Wild-type mice on GF background were bred and maintained at the Germ-free Animal Core Facility of the University of Michigan. GF mice were maintained in flexible film isolators and were checked weekly for germ-free status by aerobic and anaerobic culture. The absence of microbiota was verified by microscopic analysis of stained cecal contents to detect unculturable contamination. All animal studies were performed according to approved protocols by the University of Michigan Committee on the Use and Care of Animals. Reconstitution of GF mice with cecal contents and defined bacteria Cecal contents from single neonatal and 7-week old SPF mice were diluted into 50 ml (per one adult mouse) or 5 ml (per one 4, 12, 16-day old mouse) of PBS, and passed through a 70 μm cell strainer to eliminate clumps and debris under sterile conditions) under an atmosphere of 5% H2, 5% CO2 and 90% N2 in an anaerobic growth chamber (Coy Manufacturing, Grass Lake, MI). Then 200 μl of the suspension was processed by intragastric gavage into each of the recipient GF mice. Bacteroides species were anaerobically grown at 37°C overnight in pre-reduced chopped-meat broth (Anaerobe systems, Morgan Hill, CA). E. coli were grown aerobically overnight in LB broth with shaking. |
Treatment Protocol Filename: | Treatment_Protocol.pdf |
Treatment: | Biotic |
Treatment Compound: | Suspension of diluted cecal contents |
Treatment Route: | Intragastric gavage |
Treatment Dose: | 200 μl |