Rapid purification of RNA secondary structures.
A new method for rapid purification and structural analysis of oligoribonucleotides of 19 and 20 nt is applied to RNA hairpins SL3 and SL2, which are stable secondary structures present on the psi recognition element of HIV-1. This approach uses ion-pairing reversed-phase liquid chromatography (IP-RPLC) to achieve the separation of the stem-loop from the transcription mix. Evidence is presented that IP-RPLC is sensitive to the different conformers of these secondary structures. The purity of each stem-loop was confirmed by mass spectrometry and PAGE. IP-RPLC purification was found to be superior to PAGE in terms of time, safety and, most importantly, purity.
Department of Chemistry and Biochemistry, University of Maryland, Baltimore County, Baltimore, MD 21250, USA.
Stacy L
SL
Gelhaus
William R
WR
LaCourse
Nathan A
NA
Hagan
Gaya K
GK
Amarasinghe
Daniele
D
Fabris
D001483:Base Sequence; D002853:Chromatography, Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D015497:HIV-1 / Q000235:genetics*; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020038:RNA, Spliced Leader / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D038621:Regulatory Sequences, Ribonucleic Acid / Q000235:genetics*; D021241:Spectrometry, Mass, Electrospray Ionization; D013997:Time Factors
D009843:Oligoribonucleotides; D020038:RNA, Spliced Leader; D012367:RNA, Viral; D038621:Regulatory Sequences, Ribonucleic Acid