E very sensitive oligonucleotides can be conveniently isolated. If capping is carried out using cap A containing phenoxyacetic anhydride, it is possible to deprotect ultramIlD oligonucleotides
in 4 hours at Rt with 0.05m potassium carbonate in methanol or 2 hours at Rt with ammonium hydroxide. Alternatively, using the regular cap A containing acetic anhydride, it is necessary to deprotect overnight at room temperature to remove any Ac-dG formed during the capping step. for tAmRA containing oligonucleotides, an alternative deprotection3 may be carried out using t-butylamine/methanol/water (1:1:2) overnight at 55. Another option that we have found to be excellent uses t-butylamine/water (1:3) for 6 hours at 60. In this case, the regular protecting groups on the monomers may be used. An even milder approach has been described as “ultraultramild”.4 In this technique, Q-supports5 are combined with ultramild monomers to allow extremely gentle deprotection. After completion of the synthesis, the solid support is dried and treated overnight at 55 with a solution containing 10% (v/v) diisopropylamine (iPr2NH) in 0.25 m mercaptoethanol in meoH.
summary successful oligonucleotide cleavage and deprotection require consideration of the deprotection conditions for each product and some products may require pretreatment or special deprotection conditions. each synthesis should be reviewed to ensure the products have compatible deprotection conditions. special deprotection requirements can be found on our Analytical Reports, certificates of Analysis, technical Bulletins, and Website: http://glenresearch.
(1) S.L. Beaucage and R.P. Iyer, Tetrahedron, 1992, 48, 2223-2311. (2) M.P. Reddy, N.B. Hanna, and F. Farooqui, Nucleos Nucleot, 1997, 16, 1589-1598. (3) B. Mullah and A. Andrus, Tetrahedron Lett, 1997, 38, 5751-5754. (4) L.C.J. Gillet, J. Alzeer, and O.D. Scharer, Nucleic Acid Res, 2005, 33, 1961-1969. (5) R.T. Pon, and S.Y. Yu, Nucleic Acids Res, 1997, 25, 3629-3635.
References:
3
DEprotEction – VolumE 2 – rnA DEprotEction
IntroductIon In the previous article in this series on Deprotection (Glen Report 20.2), we focused on the absolute necessity to “Deprotect to completion” while following our mandate to “Do No Harm”.71441-28-6 manufacturer In this article, we focus our attention on RNA Deprotection.160743-62-4 Synonym Again, this is not a comprehensive review of the topic.PMID:30422520 Rather, we are attempting to offer a unified deprotection strategy that is simple to follow for newcomers to the mysterious art of RNA synthesis, while producing pure, active RNA oligos with the minimum of fuss. Where appropriate, we will mention other suitable techniques but by reference only. In the meantime, our detailed technical bulletins for RNA deprotection have also been updated and can be found on our web site by following these links:
http://glenresearch/Technical/ TB_RNA_TBDMS_Deprotection.pdf http://glenresearch/Technical/ TB_RNA_TOM_Deprotection.pdf
using methylamine. TBDMS-Protected RNA Phosphoramidites these are our workhorse monomers with an excellent cost / performance ratio. they are also compatible with high speed deprotection techniques using methylamine. UltraMild RNA Phosphoramidites many minor RNA monomers, modifiers and dyes are not compatible with aggressive deprotection techniques and these ultramild monomers will allow much milder deprotection conditions. Any downstream purification requirements will also impact the proper handling of the RNA throughout the deprotection process. for example, Dmt-on puri.MedChemExpress (MCE) offers a wide range of high-quality research chemicals and biochemicals (novel life-science reagents, reference compounds and natural compounds) for scientific use. We have professionally experienced and friendly staff to meet your needs. We are a competent and trustworthy partner for your research and scientific projects.Related websites: https://www.medchemexpress.com