Visualizing the spatiotemporal organization of the genome will improve our understanding of how chromatin structure and function are intertwined. Here, we describe the further development of the RNA-guided endonuclease-in situ labeling (RGEN-ISL) method CRISPR-FISH. Using soybean and mouse chromosomes, we demonstrate that the treatment of conventionally fixed chromosomes (in ethanol or methanol:acetic acid) with 40 mM Tris-HCl (pH 9.0) for 30 minutes at 37°C prior to CRISPR-FISH allows the application of this method for the detection of high-copy sequences. Wheat, rye, maize, and Nicotiana benthamiana were used to confirm the applicability of the identified CRISPR-FISH conditions also in other species.

中文翻译：

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Tris-HCl 的应用允许通过 CRISPR-FISH 对常规制备的染色体进行特异性标记


可视化基因组的时空组织将提高我们对染色质结构和功能如何交织在一起的理解。在这里，我们描述了RNA引导核酸内切酶原位标记（RGEN-ISL）方法CRISPR-FISH的进一步发展。使用大豆和小鼠染色体，我们证明在 CRISPR-FISH 之前，用 40 mM Tris-HCl (pH 9.0) 在 37°C 下处理常规固定染色体（在乙醇或甲醇:乙酸中） 30 分钟，可以应用该方法用于检测高拷贝序列。小麦、黑麦、玉米和本塞姆氏烟草被用来确认已识别的 CRISPR-FISH 条件在其他物种中的适用性。