Here we present a protocol for generating transgenic embryos in Xenopus laevis and Xenopus tropicalis. The method includes three steps: (1) The preparation of high-speed egg extracts, which facilitates the replacement of protamines in sperm nuclei with nucleosomes and decondenses the chromatin of sperm nuclei; (2) The isolation of sperm nuclei; and (3) The mixing of sperm nuclei, restriction enzyme, and high-speed extract in vitro, following by nuclear transplantation into unfertilized eggs to generate the transgenic embryos. This procedure generates non-mosaic transgenic embryos at high frequency and efficiency. © 2012 Springer Science+Business Media, LLC.
CITATION STYLE
Ishibashi, S., Kroll, K. L., & Amaya, E. (2012). Generating transgenic frog embryos by Restriction Enzyme Mediated Integration (REMI). Methods in Molecular Biology, 917, 185–203. https://doi.org/10.1007/978-1-61779-992-1_11
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