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Imaging rRNA Methylation in Bacteria by MR-FISH

Research output: Chapter in Book/Report/Conference proceedingChapter in a book

  • Martin R Challand
Original languageEnglish
Title of host publicationImaging Gene Expression
Publisher or commissioning bodyHumana Press
Pages89-107
Number of pages18
ISBN (Electronic)978-1-4939-9674-2
ISBN (Print)978-1-4939-9673-5
DOIs
DatePublished - 13 Aug 2019

Publication series

NameMethods in Molecular Biology

Abstract

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

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