Applications of PLOR in labeling large RNAs at specific sites.

Applications of PLOR in labeling large RNAs at specific sites.
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DOI:
10.1016/j.ymeth.2016.03.014
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发表时间:
2016-07
期刊:
影响因子:
4.8
通讯作者:
Yu Liu;P. Yu;M. Dyba;R. Sousa;J. Stagno;Yun-Xing Wang
Yu Liu;P. Yu;M. Dyba;R. Sousa;J. Stagno;Yun-Xing Wang
中科院分区:
生物学3区
文献类型:
--
作者:
Yu Liu;P. Yu;M. Dyba;R. Sousa;J. Stagno;Yun-Xing Wang

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在rna的特定位点加入修饰或标记的核苷酸对于深入了解rna的结构和功能至关重要。使用当前的方法制备适合于结构或功能研究的位点特异性标记的大rna是非常困难的。RNA的位置选择性标记,PLOR,是最近发展起来的一种使这种合成成为可能的方法。PLOR允许结合各种探针,包括2d /13C/ 15n同位素标签,Cy3/Cy5/Alexa488/Alexa555荧光染料,生物素和其他化学基团,进入长rna的特定位置。在这里,我们详细描述了PLOR在特定片段或离散位点标记rna的使用。
Incorporation of modified or labeled nucleotides at specific sites in RNAs is critical for gaining insights into the structure and function of RNAs. Preparation of site-specifically labeled large RNAs in amounts suitable for structural or functional studies is extremely difficult using current methodologies. The position-selective labeling of RNA, PLOR, is a recently developed method that makes such syntheses possible. PLOR allows incorporation of various probes, including2D/13C/15N-isotopic labels, Cy3/Cy5/Alexa488/Alexa555 fluorescent dyes, biotin and other chemical groups, into specific positions in long RNAs. Here, we describe in detail the use of PLOR to label RNAs at specific segment(s) or discrete sites.