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体外RNA的合成与标记

Synthesis and labeling of RNA in vitro.

作者信息

Huang Chao, Yu Yi-Tao

机构信息

Process Science Downstream, Bristol-Myers Squibb Company, East Syracuse, New York, USA.

出版信息

Curr Protoc Mol Biol. 2013 Apr;Chapter 4:Unit4.15. doi: 10.1002/0471142727.mb0415s102.

DOI:10.1002/0471142727.mb0415s102
PMID:23547015
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3736555/
Abstract

This unit discusses several methods for generating large amounts of uniformly labeled, end-labeled, and site-specifically labeled RNAs in vitro. The methods involve a number of experimental procedures, including RNA transcription, 5' dephosphorylation and rephosphorylation, 3' terminal nucleotide addition (via ligation), site-specific RNase H cleavage directed by 2'-O-methyl RNA-DNA chimeras, and 2-piece splint ligation. The applications of these RNA radiolabeling approaches are also discussed.

摘要

本单元讨论了几种在体外生成大量均匀标记、末端标记和位点特异性标记RNA的方法。这些方法涉及许多实验步骤，包括RNA转录、5'去磷酸化和再磷酸化、3'末端核苷酸添加（通过连接）、由2'-O-甲基RNA-DNA嵌合体指导的位点特异性RNase H切割以及两段式夹板连接。还讨论了这些RNA放射性标记方法的应用。

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