Stable Isotope Probing Techniques and Methodological Considerations Using 15N
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- 15N, Amplicon sequencing, BNF, DNA-SIP, Diazotrophs, Nitrogen, RNA-SIP,
- MeSH
- RNA, Bacterial chemistry genetics isolation & purification metabolism MeSH
- Nitrogen-Fixing Bacteria genetics metabolism MeSH
- Centrifugation, Density Gradient MeSH
- DNA, Bacterial chemistry genetics isolation & purification metabolism MeSH
- Nitrogen Fixation genetics physiology MeSH
- Isotope Labeling methods MeSH
- Nitrogen Isotopes metabolism MeSH
- High-Throughput Nucleotide Sequencing MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- RNA, Bacterial MeSH
- DNA, Bacterial MeSH
- Nitrogen Isotopes MeSH
- Nitrogen-15 MeSH Browser
Nitrogen fixation and assimilation processes are vital to the functioning of any ecosystem. Nevertheless, studying these processes using 15N-based stable isotope probing was so far limited because of technical challenges related to the relative rarity of nitrogen in nucleic acids and proteins compared to carbon, and because of its absence in lipids. However, the recent adoption of high-throughput sequencing and statistical modelling methods to SIP studies increased the sensitivity of the method and enabled overcoming some of the challenges. This chapter describes in detail how to perform DNA- and RNA-SIP using 15N.
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