R‐loops represent an abundant class of large non‐B DNA structures in genomes. Even though they form transiently and at modest frequencies, interfering with R‐loop formation or dissolution has significant impacts on genome stability. Addressing the mechanism(s) of R‐loop‐mediated genome destabilization requires a precise characterization of their distribution in genomes. A number of independent methods have been developed to visualize and map R‐loops, but their results are at times discordant, leading to confusion. Here, we review the main existing methodologies for R‐loop mapping and assess their limitations as well as the robustness of existing datasets. We offer a set of best practices to improve the reproducibility of maps, hoping that such guidelines could be useful for authors and referees alike. Finally, we propose a possible resolution for the apparent contradictions in R‐loop mapping outcomes between antibody‐based and RNase H1‐based mapping approaches.

中文翻译：

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R 循环的可视化、映射和操作的最佳实践

R 环代表基因组中丰富的一类大型非 B DNA 结构。即使它们以适度的频率瞬时形成，干扰 R 环的形成或溶解也会对基因组稳定性产生重大影响。解决 R 环介导的基因组不稳定的机制需要精确表征它们在基因组中的分布。已经开发了许多独立的方法来可视化和映射 R 循环，但它们的结果有时不一致，导致混乱。在这里，我们回顾了现有的主要 R-loop 映射方法，并评估了它们的局限性以及现有数据集的稳健性。我们提供了一组最佳实践来提高地图的可重复性，希望这些指南对作者和审稿人都有用。最后，