Introduction
The surfaces of the human body are home to microbial communities. The vast majority of the microbial cells are residing in the gastrointestinal (GI) tract. Although this microbiota is known to have influence in human health and disease, very little is known about what are they doing in the human GI tract. This is especially true for the small intestine, which is largely unexplored due to its limited accessibility. Ilesostomy subjects, people who have had their colon surgically removed because of inflammation or disease, allow access to small intestinal content in a noninvasive way and hence, can serve as a model for studying the microbiota in the small intestine. The first studies on these ileostomy subjects have shown that the microbial community species composition and function are completely different from those in the colon studies have demonstrated. To get a more complete understanding of the small intestinal bacteria and their function, we use a novel RNA sequencing tool to study microbial transcripts that reflect their in situ expressed genes in the small intestine.
This will ultimately lead to determine which microbe is performing what function in the small intestine.
Aim
The aim of this project is to determine the gene expression pattern of the small intestinal microbiota in different ileostomy subjects at different sampling points by transcriptome sequencing.
Techniques and Methods
- RNA extraction and purification
- mRNA enrichment
- PCR and agarose gel electrophoresis
- cDNA synthesis
- Sequence analysis |
