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Chapter 6 : Methods for Cyclic Di-GMP Detection
The first decade of the 21st century has seen an explosion of interest in the bacterial second messenger signal cyclic di-GMP (c-di-GMP). The study of c-di-GMP during this period has been fueled by the development of methods to visualize and quantify this molecule. This chapter reviews the history of the development of methods for c-di-GMP detection, summarizes the currently available technologies, and highlights the new challenges that must be overcome to deepen one's understanding of this molecule and its actions. The methods to measure and quantify c-di-GMP from living cells have sprung forth from the early work of the Benziman laboratory. Two major approaches, two-dimensional thin-layer chromatography (2D-TLC) and HPLC-MS-MS, currently dominate the approaches to quantify the intracellular levels of c-di-GMP, and it is likely that these two techniques will remain important tools in the toolboxes of researchers in this field. However, to gain a further understanding of this signaling molecule, new technologies measuring c-di-GMP at the single-cell level and in localized regions of the cells must be developed. The development of the new technologies will ensure the continuing renaissance in the study of c-di-GMP signaling.