TY - JOUR
T1 - Rapid purification of RNA secondary structures
AU - Gelhaus, Stacy L.
AU - LaCourse, William R.
AU - Hagan, Nathan A.
AU - Amarasinghe, Gaya K.
AU - Fabris, Daniele
N1 - Funding Information:
We would like to thank Dr Michael Summers, HHMI, for donating the stem–loop, SL2, for the project. We are deeply grateful for the financial support from Transgenomic Inc., San Jose, CA. The SL3 synthesis and MS analysis was supported by NIH Grant R01-GM643208 (D.F.).
PY - 2003/11/1
Y1 - 2003/11/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0642365197&partnerID=8YFLogxK
U2 - 10.1093/nar/gng136
DO - 10.1093/nar/gng136
M3 - Article
C2 - 14576335
AN - SCOPUS:0642365197
SN - 0305-1048
VL - 31
SP - e135
JO - Nucleic acids research
JF - Nucleic acids research
IS - 21
ER -