RNA-Based Stable Isotope Probing (RNA-SIP) in the Gut Environment

Methods Mol Biol. 2019:2046:221-231. doi: 10.1007/978-1-4939-9721-3_17.

Abstract

The RNA-SIP technology allows for linking the structure and function of complex microbial communities, that is, the identification of microbial key players involved in distinct degradation and assimilation processes under in situ conditions. Being dependent on RNA, this technique is particularly suited for environments with high numbers of very active, that is, significantly RNA-expressing microorganisms, such as intestinal tract samples. We use RNA-SIP for the identification of bacteria involved in the degradation and assimilation of prebiotic carbohydrates in order to better understand the functionality of these medically and economically important nutrients in human and animal intestinal environments.

Keywords: Digestion; Glucose; Intestinal tract; Prebiotics; RNA-SIP; Resistant starch.

MeSH terms

  • Animals
  • Bacteria / genetics
  • Bacteria / metabolism
  • Carbon Isotopes / metabolism*
  • Centrifugation, Density Gradient
  • Glucose / metabolism
  • Humans
  • Intestines / microbiology*
  • Isotope Labeling / methods*
  • Microbiota / genetics
  • Microbiota / physiology
  • Prebiotics / microbiology
  • RNA Probes / metabolism
  • RNA, Bacterial / isolation & purification
  • RNA, Bacterial / metabolism*
  • Starch / metabolism

Substances

  • Carbon Isotopes
  • Prebiotics
  • RNA Probes
  • RNA, Bacterial
  • Starch
  • Glucose