Synthesis of backbone deuterium labelled [r(CGCGAAUUCGCG)]2 and HPLC purification of synthetic RNA
- PMID: 1383940
- PMCID: PMC334295
- DOI: 10.1093/nar/20.19.5131
Synthesis of backbone deuterium labelled [r(CGCGAAUUCGCG)]2 and HPLC purification of synthetic RNA
Abstract
The chemical synthesis of backbone deuterium labelled [r(CGCGAAU*U*CGCG)]2 (U* = [5'-2H]U) is described. An efficient purification procedure was developed using a polymeric reverse phase (PRP) HPLC column at 60 degrees C. This procedure provided pure RNA dodecamer in the multi-milligram quantities (39% overall yield) necessary for dynamics studies using solid-state deuterium NMR. The purification method has been effectively applied to other RNA sequences and will assist biophysical studies which require relatively large quantities of RNA oligomers.
Similar articles
-
A general purification procedure for chemically synthesized oligoribonucleotides.Anal Biochem. 1994 Apr;218(1):177-84. doi: 10.1006/abio.1994.1157. Anal Biochem. 1994. PMID: 7519835
-
Advances in the chemical synthesis and purification of RNA.Nucleic Acids Symp Ser. 1995;(33):123-5. Nucleic Acids Symp Ser. 1995. PMID: 8643345
-
Solid-phase synthesis and high-resolution NMR studies of two synthetic double-helical RNA dodecamers: r(CGCGAAUUCGCG) and r(CGCGUAUACGCG).Biochemistry. 1989 Mar 21;28(6):2422-35. doi: 10.1021/bi00432a013. Biochemistry. 1989. PMID: 2471547
-
Isotope labeling and segmental labeling of larger RNAs for NMR structural studies.Adv Exp Med Biol. 2012;992:121-44. doi: 10.1007/978-94-007-4954-2_7. Adv Exp Med Biol. 2012. PMID: 23076582 Review.
-
Chemo-enzymatic labeling for rapid assignment of RNA molecules.Methods. 2016 Jul 1;103:11-7. doi: 10.1016/j.ymeth.2016.03.015. Epub 2016 Apr 18. Methods. 2016. PMID: 27090003 Review.
Cited by
-
Stabilization of RNA stacking by pseudouridine.Nucleic Acids Res. 1995 Dec 25;23(24):5020-6. doi: 10.1093/nar/23.24.5020. Nucleic Acids Res. 1995. PMID: 8559660 Free PMC article.
-
HPLC purification of RNA for crystallography and NMR.RNA. 1996 Feb;2(2):110-7. RNA. 1996. PMID: 8601278 Free PMC article.
-
Synthesis and studies on the effect of 2-thiouridine and 4-thiouridine on sugar conformation and RNA duplex stability.Nucleic Acids Res. 1997 Mar 15;25(6):1272-80. doi: 10.1093/nar/25.6.1272. Nucleic Acids Res. 1997. PMID: 9092639 Free PMC article.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials