电子显微镜下染色质的基因特异性标记

John D. Brantley , Michael Beer
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引用次数: 1

摘要

在限制性内切酶酶解的非洲爪蟾红细胞染色质中,用胶体金特异性标记5S卵母细胞基因,以便在电镜下进行鉴定。核首先用HindIII限制性内切酶酶切,然后用T7酶切,在片段上留下单链3 '尾。将5S基因3 '端与尾部互补的生物素化15-mer寡核苷酸退火到染色质上,链霉亲和素络合胶体金与之结合。在电子显微镜的扩散中,标记的片段很容易被识别。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Gene-specific labeling of chromatin for electron microscopy

In restriction enzyme digested Xenopus laevis erythrocyte chromatin, the 5S oocyte genes were specifically labeled with colloidal gold so they could be identified in an electron microscopic study. Nuclei were digested first with HindIII restriction endonuclease followed by T7 exonuclease to leave single-stranded 3′ tails on the fragments. A biotinylated 15-mer oligonucleotide complementary to the tail on the 3′ end of the 5S gene was annealed to the chromatin, and streptavidin complexed colloidal gold was bound to it. In a spread for electron microscopy the labeled fragments were readily recognized.

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