RNA analysis by ion-pair reversed-phase high performance liquid chromatography
RNA analysis by ion-pair reversed-phase high performance liquid chromatography
复制标题
DOI：
10.1093/nar/29.2.e7
复制
发表时间：
2001-01-15
影响因子：
14.9
通讯作者：
Hecker, Karl H.
中科院分区：
文献类型：
--
作者：
Azarani, Arezou;Hecker, Karl H.
离子对反相高效液相色谱（iprp - HPLC）是一种新的、优越的RNA分析方法。IP RP高效液相色谱法提供了一种快速、可靠的替代传统的RNA分析方法，包括不同RNA种类的分离、定量和纯化。RNA在所用的分析条件下是稳定的；在分析过程中没有观察到RNA的降解。证明了IP - R高效液相色谱用于RNA分析的通用性。RNA阶梯的组成部分，大小范围从155到1770 nt，被分解。分析了多达5219nt的RNA转录本，确定了它们的完整性，并对它们进行了定量和纯化。描述了从总RNA中纯化mRNA，从poly(A)(+) mRNA中分离小鼠rRNA。IP RP高效液相色谱法也适用于digi标记RNA和未标记RNA的分离纯化。用IP RP高效液相色谱法纯化的RNA具有更好的稳定性。
Ion-pair reversed-phase high performance liquid chromatography (IP RP HPLC) is presented as a new, superior method for the analysis of RNA. IP RP HPLC provides a fast and reliable alternative to classical methods of RNA analysis, including separation of different RNA species, quantification and purification. RNA is stable under the analysis conditions used; degradation of RNA during the analyses was not observed. The versatility of IP R HPLC for RNA analysis is demonstrated. Components of an RNA ladder, ranging in size from 155 to 1770 nt, were resolved. RNA transcripts of up to 5219 nt were analyzed, their integrity determined and they were quantified and purified. Purification of mRNA from total RNA is described, separating mouse rRNA from poly(A)(+) mRNA. IP RP HPLC is also suitable for the separation and purification of DIG-labeled from unlabeled RNA. RNA purified by IP RP HPLC exhibits improved stability.