2019-08-13
Imaging rRNA methylation in bacteria by MR-FISH
Publication
Publication
Methods Mol. Biol. , Volume 2038 p. 89- 107
Methylation of RNA is normally monitored in purified cell lysates using next-generation sequencing, gel electrophoresis, or mass spectrometry as readouts. These bulk methods require the RNA from ~104 to 107 cells to be pooled to generate sufficient material for analysis. Here we describe a method—methylation-sensitive RNA in situ hybridization (MR-FISH)—that assays rRNA methylation in bacteria on a cell-by-cell basis, using methylation-sensitive hybridization probes and fluorescence microscopy. We outline step-by-step protocols for designing probes, in situ hybridization, and analysis of data using freely available code.
| Additional Metadata | |
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| Springer Nature | |
| doi.org/10.1007/978-1-4939-9674-2_7 | |
| Methods Mol. Biol. | |
| Organisation | Physics of Cellular Interactions |
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Ganzinger, K., Challand, M., Spencer, J., Klenerman, D., & Ranasinghe, R. (2019). Imaging rRNA methylation in bacteria by MR-FISH. In Methods Mol. Biol. (Vol. 2038, pp. 89–107). doi:10.1007/978-1-4939-9674-2_7 |
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