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Microbial Diversity in Intestinal Regions of WT Mice: A Comparative Pyrosequencing Analysis

This study investigates the microbial composition in the interfold and digesta regions of WT mice using histology, LCM, DNA extraction, pyrosequencing, and PCR assays. The analysis includes random cloning, Sanger sequencing, TRFLP, and multivariate techniques to identify bacterial families like Bacteroidaceae, Enterococcaceae, Lactobacillaceae, and Lachnospiraceae-Ruminococcaceae. Known type strains and phylogenetic analysis provide insights into the microbial diversity observed in different regions. Results show significant differences in microbial density and composition between interfold and digesta regions.

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Microbial Diversity in Intestinal Regions of WT Mice: A Comparative Pyrosequencing Analysis

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  1. Sup. Fig. 1 Sample set #1 WT mice (n = 3) Sample set #2 WT mice (n = 3) Sample set #3 WT mice (n = 3) Histology LCM of microbes DNA extraction Histology LCM of microbes DNA extraction Histology LCM of microbes DNA extraction Pyrosequencing analysis Interfold region (29,560 reads) Digesta region (38,120 reads) • Microbial density by • quantitative PCR assays • Interfold vs. digesta bacterial families: • Bacteroidaceae* • Enterococcaceae • Lactobacillaceae • Lachnospiraceae-Ruminococcaceae Random cloning and Sanger sequencing Interfold region (496 clones) Digesta region (377 clones) • TRFLP and multivariate analysis • Interfold vs. digesta • Lachnospiraceae-Ruminococcaceae • 16S rRNA gene classification and phylogenetic analysis • Known type strains¥ • Lachnospiraceae-Ruminococcaceae

  2. Sup. Fig. 2 A B

  3. Sup. Fig. 3 A B C

  4. Sup. Fig. 4 Interfold Digesta 97% 97% 95% 95% Number of OTUs observed 90% 90% 80% 80% Number of sequences

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