Dinchuk J E, Kelley K, Boyle A L
Miles, Inc., West Haven, CT.
Biotechniques. 1994 Nov;17(5):954-61.
Homozygosity in transgenic mice is typically confirmed by the time-consuming practice of breeding suspected homozygotes to non-transgenics and looking for 100% transmittance of the transgene to offspring. We have devised a simple reproducible method for isolating white blood cell nuclei from small quantities of mouse whole blood and have utilized fluorescence in situ hybridization to confirm the presence of one (hemizygous) or two (homozygous) copies of the transgene locus in interphase nuclei fixed to microscope slides. This method should have other applications where in situ hybridization of interphase nuclei is desirable.
转基因小鼠的纯合性通常通过将疑似纯合子与非转基因小鼠交配这一耗时的方法来确认,并观察转基因向后代的100%传递情况。我们设计了一种简单且可重复的方法,用于从少量小鼠全血中分离白细胞核,并利用荧光原位杂交技术来确认固定在显微镜载玻片上的间期核中转基因位点的一个(半合子)或两个(纯合子)拷贝的存在。该方法在需要进行间期核原位杂交的其他应用中也应具有适用性。