RNA analysis by ion-pair reversed-phase high performance liquid chromatography

被引：56

作者：

Azarani, Arezou ^{[1
]}

Hecker, Karl H. ^{[1
]}

机构：

[1] Transgenomic Inc, San Jose, CA 95131 USA

来源：

NUCLEIC ACIDS RESEARCH | 2001年 / 29卷 / 02期

关键词：

D O I：

10.1093/nar/29.2.e7

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

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页数：9

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