使用 GRO-seq 对小麦中新生 RNA 进行经济高效的分析方案。
Protocol for affordable and efficient profiling of nascent RNAs in bread wheat using GRO-seq.
机构信息
Guangdong Provincial Key Laboratory of Plant Adaptation and Molecular Design, Guangzhou Key Laboratory of Crop Gene Editing, Innovative Center of Molecular Genetics and Evolution, School of Life Sciences, Guangzhou University, Guangzhou 510006, China.
National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
出版信息
STAR Protoc. 2022 Sep 16;3(3):101657. doi: 10.1016/j.xpro.2022.101657. Epub 2022 Sep 2.
Exorbitant sequencing cost is one of the main obstacles limiting the widespread application of Global Run-On sequencing (GRO-seq) to detect transcriptional activity. Here, we describe a more efficient and affordable protocol for GRO-seq that incorporates an rRNA removal step after nuclear RNA isolation and before nascent RNA immunoprecipitation. We have successfully applied this protocol to profile enhancer transcription in allohexaploid bread wheat and increased the proportion of valid data by 20 times. For complete details on the use and execution of this protocol, please refer to Xie et al. (2022).
测序成本过高是限制全球运行测序（GRO-seq）广泛应用于检测转录活性的主要障碍之一。在这里，我们描述了一种更有效和经济的 GRO-seq 方案，该方案在核 RNA 分离后和新生 RNA 免疫沉淀前加入 rRNA 去除步骤。我们已经成功地将该方案应用于异源六倍体面包小麦的增强子转录谱分析，并将有效数据的比例提高了 20 倍。有关该方案的使用和执行的完整详细信息，请参考 Xie 等人（2022 年）。
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