一种用于RNA 3'端标记的简单方法。
A simple method for 3'-labeling of RNA.
作者信息
Huang Z, Szostak J W
机构信息
Department of Molecular Biology, Massachusetts General Hospital, Boston 02114, USA.
出版信息
Nucleic Acids Res. 1996 Nov 1;24(21):4360-1. doi: 10.1093/nar/24.21.4360.
Abstract
We describe a simple method for 3'-end labeling RNAs of known sequence. A short DNA template is designed to anneal to the 3'-end of the RNA, with a two nucleotide 5' overhang of 3'-TA-5', 3'-TG-5' or 3'-TC-5'. The Klenow fragment of DNA polymerase I can then cleanly and efficiently extend the 3'-end of the RNA by the incorporation of a single alpha-32P-labeled dATP residue. This method can be used to label one RNA in a mixture of RNAs, or to label 5'-blocked RNAs such as mRNA.
摘要
我们描述了一种用于对已知序列的RNA进行3'末端标记的简单方法。设计一个短DNA模板，使其与RNA的3'末端退火，具有3'-TA-5'、3'-TG-5'或3'-TC-5'的两个核苷酸5'突出端。然后，DNA聚合酶I的Klenow片段可以通过掺入单个α-32P标记的dATP残基，干净利落地有效地延伸RNA的3'末端。该方法可用于标记RNA混合物中的一种RNA，或标记5'端封闭的RNA，如mRNA。
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