首页> 外文期刊>Analytical Biochemistry: An International Journal of Analytical and Preparative Methods >Whole-mount in situ detection of microRNAs on Arabidopsis tissues using Zip Nucleic Acid probes
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Whole-mount in situ detection of microRNAs on Arabidopsis tissues using Zip Nucleic Acid probes

机译:使用Zip核酸探针对拟南芥组织上的microRNA进行整装原位检测

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MicroRNAs (miRNAs) affect fundamental processes of development. In plants miRNAs regulate organ development, transition to flowering, and responses to abiotic/biotic stresses. To understand the biological role of miRNAs, in addition to identifying their targeted transcripts, it is necessary to characterize the spatiotemporal regulation of their expression. Many methods have been used to define the set of organ-specific miRNAs by tissue dissection and miRNA profiling but none of them can describe their tissue and cellular distribution at the high resolution provided by in situ hybridization (ISH). This article describes the setup and optimization of a whole-mount ISH protocol to target endogenous miRNAs on intact Arabidopsis seedlings using DIG-labeled Zip Nucleic Acid (ZNA) oligonucleotide probes. Automation of the main steps of the procedure by robotized liquid handling has also been implemented in the protocol for best reproducibility of results, enabling running of ISH experiments at high throughput.
机译:微小RNA(miRNA)影响基本的发育过程。在植物中,miRNA调节器官发育,过渡至开花以及对非生物/生物胁迫的反应。要了解miRNA的生物学作用,除了鉴定其目标转录物外,还必须表征其表达的时空调控。许多方法已用于通过组织解剖和miRNA分析来定义器官特异性miRNA的集合,但它们均无法以原位杂交(ISH)提供的高分辨率描述其组织和细胞分布。本文介绍了使用DIG标记的Zip核酸(ZNA)寡核苷酸探针针对完整的拟南芥幼苗上靶向内源性miRNA的完整ISH协议的设置和优化。协议中还通过自动液体处理实现了流程主要步骤的自动化,以实现结果的最佳重现性,从而能够以高通量运行ISH实验。

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