AI Article Synopsis
- Stable isotope probing (SIP) is a technique that helps identify and study the roles of uncultivated microorganisms in environmental samples by using stable isotopes.
- RNA-SIP specifically leverages the increase in RNA density when heavier isotopes like carbon and nitrogen are taken up by active microbes, allowing for separation of labeled RNA for analysis.
- The chapter outlines a detailed protocol for conducting RNA-SIP experiments, covering steps such as extraction, ultracentrifugation, and analyzing labeled RNAs.
Article Abstract
Stable isotope probing is a combined molecular and isotopic technique used to probe the identity and function of uncultivated microorganisms within environmental samples. Employing stable isotopes of common elements such as carbon and nitrogen, RNA-SIP exploits an increase in the buoyant density of RNA caused by the active metabolism and incorporation of heavier mass isotopes into the RNA after cellular utilization of labeled substrates pulsed into the community. Labeled RNAs are subsequently separated from unlabeled RNAs by density gradient centrifugation followed by identification of the RNAs by sequencing. Therefore, RNA stable isotope probing is a culture-independent technique that provides simultaneous information about microbiome community, composition and function. This chapter presents the detailed protocol for performing an RNA-SIP experiment, including the formation, ultracentrifugation, and fractional analyses of stable isotope-labeled RNAs extracted from environmental samples.
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| http://dx.doi.org/10.1007/978-1-4939-9721-3_3 | DOI Listing |
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