A simplified and rapid in situ hybridization protocol for planarians
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Description
Whole-mount in situ hybridization is a critical technique for analyzing gene expression in planarians. While robust in situ protocols have been developed, these protocols are laborious, making them challenging to incorporate in an academic setting, reducing throughput and increasing time to results. Here, the authors systematically tested modifications to all phases of the protocol with the goal of eliminating steps and reducing time without impacting quality. This modified protocol allows for whole-mount colorimetric in situ hybridization and multicolor fluorescence in situ hybridization to be completed in two days with a significant reduction in steps and hands-on processing time.
Publication Date
10-18-2023
Keywords
antibody blocking • bromodeoxyuridine • Dugesia japonica • fixation • gene expression analysis • immunofluorescence • in situ hybridization • planarians • Schmidtea mediterranea • stem cell
Recommended Citation
Gaetano, Andrew J. and King, Ryan, "A simplified and rapid in situ hybridization protocol for planarians" (2023). Faculty Creative and Scholarly Works. 60.
https://digitalcommons.snc.edu/faculty_staff_works/60
Comments
This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/