Azarani A, Hecker K H
Transgenomic Inc., 2032 Concourse Drive, San Jose, CA 95131, USA.
Nucleic Acids Res. 2001 Jan 15;29(2):E7. doi: 10.1093/nar/29.2.e7.
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 RP 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.
离子对反相高效液相色谱法(IP RP HPLC)被提出作为一种分析RNA的全新且优越的方法。IP RP HPLC为RNA分析的传统方法提供了一种快速且可靠的替代方法,包括不同RNA种类的分离、定量和纯化。在所使用的分析条件下RNA是稳定的;在分析过程中未观察到RNA的降解。证明了IP RP HPLC在RNA分析方面的多功能性。解析了大小从155到1770 nt的RNA梯状条带的各个组分。分析了长达5219 nt的RNA转录本,确定了它们的完整性,并对其进行了定量和纯化。描述了从总RNA中纯化mRNA的方法,即将小鼠rRNA与聚腺苷酸(+)mRNA分离。IP RP HPLC也适用于从未标记的RNA中分离和纯化地高辛标记的RNA。通过IP RP HPLC纯化的RNA表现出更高的稳定性。