适用于快速凝胶测序的用32P对RNA进行3'末端标记
3'End labelling of RNA with 32P suitable for rapid gel sequencing.
作者信息
Winter G, Brownlee G G
出版信息
Nucleic Acids Res. 1978 Sep;5(9):3129-39. doi: 10.1093/nar/5.9.3129.
Abstract
A new general method of labelling the 2',3'-diol end of RNA with 32P has been devised suitable for gel sequencing. Poly(A) polymerase (E. coli) is incubated with the RNA and limiting amounts of alpha-32P-ATP. The mono-addition product is then cleaved with periodate and beta-eliminated with aniline, leaving the RNA terminally labelled with 3' 32P-phosphate. When applied to a model compound, tRNAPhe from E. coli, over 28 residues could be read from the 3' end.
摘要
已设计出一种用³²P标记RNA 2',3'-二醇末端的新通用方法，适用于凝胶测序。将聚（A）聚合酶（大肠杆菌）与RNA和限量的α-³²P-ATP一起孵育。然后用过碘酸盐切割单加成产物，并用苯胺进行β-消除，使RNA末端标记有3'³²P-磷酸。当应用于模型化合物——来自大肠杆菌的tRNAPhe时，从3'末端可以读出超过28个残基。
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