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XB-ART-45857
Methods Mol Biol January 1, 2012; 917 185-203.

Generating transgenic frog embryos by restriction enzyme mediated integration (REMI).



Abstract
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.

PubMed ID: 22956089
PMC ID: PMC3818119
Article link: Methods Mol Biol
Grant support: [+]

References [+] :
Amaya, Frog genetics: Xenopus tropicalis jumps into the future. 1998, Pubmed, Xenbase


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