Reversed-phase ion-pair liquid chromatography analysis and purification of small interfering RNA.
Small interfering RNA (siRNA)-induced gene silencing shows great promise in genomic research and therapeutic applications. siRNA duplexes are typically assembled from complementary synthetic oligonucleotides. High-purity single-stranded species are required for in vivo applications. Methods for separation, characterization, and purification of short RNA strands have been developed based on reversed-phase ion-pair liquid chromatography. The purification strategies were developed for both single-stranded and duplex RNA species. The method of duplex purification uses on-column annealing of complementary RNA strands, followed by separation of the target duplex from truncated duplexes and single-stranded RNA forms. The proposed method significantly reduces the purification time of synthetic siRNA.
Waters Corporation, Milford, MA 01757, USA. sean_m_mccarthy@waters.com
Sean M
SM
McCarthy
Martin
M
Gilar
John
J
Gebler
D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000737:chemistry; D013058:Mass Spectrometry / Q000191:economics / Q000379:methods*; D009841:Oligonucleotides / Q000032:analysis* / Q000737:chemistry; D034741:RNA, Small Interfering / Q000032:analysis* / Q000737:chemistry
D009841:Oligonucleotides; D034741:RNA, Small Interfering; D004247:DNA