Automated parallel isolation of multiple species of non-coding RNAs by the reciprocal circulating chromatography method
Automated parallel isolation of multiple species of non-coding RNAs by the reciprocal circulating chromatography method
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DOI：
10.1093/nar/gkl1129
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发表时间：
2007-02-01
影响因子：
14.9
通讯作者：
Suzuki, Tsutomu
中科院分区：
文献类型：
--
作者：
Miyauchi, Kenjyo;Ohara, Tomoya;Suzuki, Tsutomu
最近的全基因组转录组分析已经确定了不同类别的非编码RNA（ncRNA），其中一些已被证明是参与各种生物过程的功能性调节RNA。RNA分子通过各种转录后加工事件（包括两端的剪接、修饰、编辑和修剪）的成熟是RNA分子正确折叠和适当功能所需的。为了表征完全加工的天然RNA的转录后修饰和末端化学结构，有必要从有限数量和复杂的细胞RNA混合物中分离单个RNA种类。然而，还没有通用的和方便的策略来分离单个RNA。我们在这里描述的第一个例子的自动化平行分离的个人ncRNA使用一种新的方法命名为“互惠循环色谱法（RCC层析）”。RCC采用填充有固相DNA探针的多个尖端柱从总RNA的共同样品中分离多个RNA种类。中试RCC仪器成功地从E.大肠杆菌、酵母菌和小鼠。
Recent genome-wide transcriptome analysis has identified diverse classes of non-coding RNAs (ncRNAs), some of which have been demonstrated to be functional, regulatory RNAs involved in various biological processes. Maturation of RNA molecules through various post-transcriptional processing events, including splicing, modification, editing and trimming of both ends, is required for correct folding and proper function of RNA molecules. To characterize post-transcriptional modifications and terminal chemical structures of fully processed native RNAs, it is necessary to isolate individual RNA species from a limited quantity and complex mixture of cellular RNAs. However, there have been no general and convenient strategies for isolation of individual RNAs. We describe here the first example of automated parallel isolation of individual ncRNAs using a novel method named 'reciprocal circulating chromatography (RCC)'. RCC employs multiple tip-columns packed with solid-phase DNA probes to isolate multiple RNA species from a common sample of total RNAs. A pilot RCC instrument successfully isolated various ncRNAs from E. coli, yeast and mouse.