Wolff, AndrewWagner, CynthiaWolf, JuliaLobo, Daniel2024-03-062024-03-062022-06-14Wolff, Andrew, Cynthia Wagner, Julia Wolf, and Daniel Lobo. “In Situ Probe and Inhibitory RNA Synthesis Using Streamlined Gene Cloning with Gibson Assembly.” STAR Protocols 3, no. 3 (September 16, 2022): 101458. https://doi.org/10.1016/j.xpro.2022.101458.https://doi.org/10.1016/j.xpro.2022.101458http://hdl.handle.net/11603/31859The synthesis of single-stranded riboprobes or double-stranded RNAs for in situ hybridization and gene knockdowns often use vectors that require time-consuming plasmid restriction digests and inefficient gel purifications. Here, we present a faster protocol for the simultaneous plasmid restriction digestion and Gibson assembly of vectors for the synthesis of both riboprobes and double-stranded RNAs for in situ and RNA interference experiments, respectively. We illustrate the protocol with planaria in situ and RNAi assays, but it is applicable to any organism.17 pagesen-USThis item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.Creative Commons Attribution 4.0 International (CC BY 4.0)https://creativecommons.org/licenses/by/4.0/Developmental biologyModel OrganismsMolecular BiologyText