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Stool Microbial DNA Isolation: Studying the Effect of Environmental Enrichment on Microbiome Diversity in Murine Tumor Model
Overview
This video describes DNA isolation protocol from stool samples of colon tumor-bearing mice. The extracted DNA is amplified using PCR, and the product is further cleaned using a magnetic beads-based approach. This method can be used to determine the effect of environmental enrichment on colon microbiota and animal mortality.
Protocol
All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Genomic DNA Isolation from Stool
NOTE: Utilize a commercial kit to isolate microbial DNA from stool following a stool pathogen detection protocol. Remove samples directly from the -80 ˚C freezer and store on dry ice while weighing.
- Transfer up to 220 mg of stool to a clean mic.......
Representative Results
Figure 1: EE and NE housing conditions and stool homogenates (as described in the protocol)
Figure 2: 16S Microbiome Library Preparation. (A) Unpurified PCR amplicon products derived from stool genomic DNA.(B) Index.......
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Source: Fuller, A. K, et al. A Method to Define the Effects of Environmental Enrichment on Colon Microbiome Biodiversity in a Mouse Colon Tumor Model. J. Vis. Exp. (2018).
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