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非放射性病毒和信使核糖核酸的寡核苷酸图谱分析

Oligonucleotide mapping of non-radioactive virus and messenger RNAs.

作者信息

Frisby D

出版信息

Nucleic Acids Res. 1977 Sep;4(9):2975-96. doi: 10.1093/nar/4.9.2975.

Abstract

A modification of the known method for obtaining radioactive fingerprints from non-radioactive nucleic acids by labelling a digest with 5'-hydroxyl polynucleotide kinase and [gamma-32P]-ATP has been applied to RNase T1 digests from various high molecular weight virus RNAs and to ovalbumin mRNA. Fractionation of the resultant [32P]-labelled T1 RNase digests by two-dimensional polyacrylamide electrophoresis demonstrates that in the case of virus RNAs, the fingerprints thus obtained are very similar to those derived from uniformly labelled RNAs. The value of this technique is that it requires only 1-5 microgram of purified virus RNA and at least three orders of magnitude less radioactivity than is routinely employed in preparing uniformly labelled RNA.

摘要

一种通过用5'-羟基多核苷酸激酶和[γ-32P]-ATP标记消化产物从非放射性核酸获得放射性指纹图谱的已知方法的改进,已应用于各种高分子量病毒RNA的核糖核酸酶T1消化产物和卵清蛋白mRNA。通过二维聚丙烯酰胺凝胶电泳对所得的[32P]标记的T1核糖核酸酶消化产物进行分级分离表明,在病毒RNA的情况下,由此获得的指纹图谱与从均匀标记的RNA衍生的指纹图谱非常相似。该技术的价值在于,它仅需要1-5微克纯化的病毒RNA,并且所需的放射性比常规制备均匀标记RNA时使用的放射性至少少三个数量级。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0a17/342628/922e9c9985ef/nar00482-0039-a.jpg

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