期刊
ANALYTICAL BIOCHEMISTRY
卷 390, 期 2, 页码 181-188出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2009.03.042
关键词
Oligonucleotide; Duplex; Double-stranded; RNA; DNA; Ultra-performance liquid chromatography; (UPLC); HPLC; Mass spectrometry
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. (C) 2009 Elsevier Inc. All rights reserved.
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