Genome-Wide Analysis of Long Noncoding RNA Turnover
Genome-Wide Analysis of Long Noncoding RNA Turnover
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DOI：
10.1007/978-1-4939-2253-6_19
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发表时间：
2015-01-01
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影响因子：
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通讯作者：
Akimitsu, Nobuyoshi
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文献类型：
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作者：
Tani, Hidenori;Imamachi, Naoto;Akimitsu, Nobuyoshi
测定RNA周转的全基因组分析是RNA生物学中一种先进的方法，可检测核非编码RNA （ncRNA）的特定半衰期。特别是，使用尿苷类似物的脉冲标记方法能够在生理未受干扰的条件下测定RNA稳定性。该技术包括用5'-溴尿苷（BrU）对哺乳动物细胞中的内源性rna进行脉冲标记，然后使用深度测序测量BrU标记rna的时间顺序减少。这种方法被称为BrU免疫沉淀追踪法（BRIC）或BRIC through deep sequencing （BRIC-seq）。在这里，我们描述了一个详细的协议和技术提示为金砖四国序列。
Genome-wide analysis for determining RNA turnover is an advanced method in RNA biology that examines the specific half-life of nuclear noncoding RNA (ncRNA). In particular, a pulse-labeling method using uridine analogs enables the determination of RNA stability under physiologically undisturbed conditions. The technique involves pulse labeling of endogenous RNAs in mammalian cells with 5'-bromo-uridine (BrU), followed by measuring the chronological decrease of BrU-labeled RNAs using deep sequencing. The method is called BrU immunoprecipitation chase assay (BRIC) or BRIC through deep sequencing (BRIC-seq). Here, we describe a detailed protocol and technical tips for BRIC-seq.