A Proximity Ligation-Based Method to Detect RNA-DNA Association.
A Proximity Ligation-Based Method to Detect RNA-DNA Association.
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一种基于邻近连接的方法来检测 RNA-DNA 关联。
DOI：
10.1007/978-1-4939-9537-0_10
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Petruk,Svetlana
中科院分区：
文献类型：
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作者：
Fenstermaker,TylerK;Sun,Guizhi;Mazo,Alexander;Petruk,Svetlana
我们最近开发了一种使用邻近连接分析(PLA)来评估RNA-DNA相互作用的方法。这项技术被称为“RNA-DNA相互作用分析”(RDIA)，包括分别用EDU和Bru对DNA和RNA进行差异标记。一旦标记，就会进行聚乳酸以评估标记的分子是否非常接近。在这里，我们提供了利用当前商业上可用的BrdU抗体的修改的RDIA方案的详细描述。作为一个例子，我们展示了它在培养的H1299细胞和小鼠胚胎干细胞中检测最近合成的DNA上的新生转录的能力。
We recently developed a method for assessing RNA-DNA interactions using proximity ligation assays (PLA). This technique, termed the “RNA-DNA interaction assay” (RDIA), involves differentially labeling DNA and RNA with EdU and BrU, respectively. Once labeled, PLA is performed to assess if the labeled molecules are in close proximity. Here we provide a detailed description of the modified RDIA protocol utilizing currently commercially available BrdU antibodies. As an example, we show its ability to detect nascent transcripts on recently synthesized DNA in both cultured H1299 cells and mouse embryonic stem cells.