Isolation of Newly Transcribed RNA Using the Metabolic Label 4-Thiouridine.
Isolation of Newly Transcribed RNA Using the Metabolic Label 4-Thiouridine.
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使用代谢标签4-硫岛碱分离新转录的RNA。
DOI：
10.1007/978-1-4939-7204-3_13
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Hertel KJ
中科院分区：
文献类型：
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作者：
Garibaldi A;Carranza F;Hertel KJ
分离新转录的RNA是一种非常有价值的方法，可以用来研究RNA在纤维素中的动态生命。传统的全细胞RNA提取方法将后续的基因表达分析限制在RNA丰度的稳态水平，这往往掩盖了RNA合成和加工的变化。本章描述了一种低细胞毒性的方法，该方法允许标记和分离细胞培养中的新生前-mRNA。得到的分离株适合用于一系列下游应用，目的是研究RNA合成、加工或稳定性的变化。
Isolation of newly transcribed RNA is an invaluable approach that can be used to study the dynamic life of RNA in cellulo. Traditional methods of whole-cell RNA extraction limit subsequent gene expression analyses to the steady-state levels of RNA abundance, which often masks changes in RNA synthesis and processing. This chapter describes a methodology with low cytotoxicity that permits the labeling and isolation of nascent pre-mRNA in cell culture. The resulting isolate is suitable for use in a series of downstream applications aimed at studying changes in RNA synthesis, processing, or stability.