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电子显微镜下的DNA标记

DNA Labeling at Electron Microscopy.

作者信息

Thelen Nicolas, Thiry Marc

机构信息

Giga Neurosciences, Department of Life Sciences, Laboratory of Cell Biology, University of Liège, Quartier Hôpital, 15 Avenue Hippocrate (B36), 4000, Liege, Belgium.

出版信息

Methods Mol Biol. 2017;1560:269-276. doi: 10.1007/978-1-4939-6788-9_20.

Abstract

Here, we describe a method for locating DNA on ultrathin sections. This technique is compatible with all usual fixation and embedding procedures and can be combined with cytochemical methods. Ultrathin sections are incubated in a medium containing terminal deoxynucleotidyl transferase (TdT) and various non-isotopic nucleotide analogs. The labeled nucleotides bound to the surface of ultrathin sections are then visualized by an indirect immunogold labeling technique. This high-resolution method provides a powerful tool for pinpointing the precise location of DNA within biological material, even where DNA is present in very low amounts.

摘要

在此,我们描述一种在超薄切片上定位DNA的方法。该技术与所有常规固定和包埋程序兼容,并且可以与细胞化学方法相结合。超薄切片在含有末端脱氧核苷酸转移酶(TdT)和各种非同位素核苷酸类似物的培养基中孵育。然后通过间接免疫金标记技术使结合在超薄切片表面的标记核苷酸可视化。这种高分辨率方法为精确确定生物材料中DNA的位置提供了一个强大的工具,即使DNA含量非常低的情况下也是如此。

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