5′‐Iodination of Solid‐Phase‐Linked Oligodeoxyribonucleotides
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- Abstract
- Table of Contents
- Materials
- Figures
- Literature Cited
Abstract
5??Iodinated oligodeoxyribonucleotides readily react with 3??phosphorothioated DNA in the presence of a complementary template to yield a conjugate that is identical to natural DNA in every respect except that one oxygen atom in the phosphodiester backbone is replaced by a sulfur atom. The 5??iodo group is easily converted to a variety of other functional groups and will quickly react with thiol?containing labels to yield stable thioether conjugates. This unit presents manual and automated procedures for converting the 5??hydroxyl of protected CPG?bound oligodeoxyribonucleotides to an iodo group and for releasing and purifying the products.
Keywords: iodination; 5??modification; solid?phase; oligodeoxyribonucleotide; electrophile; ligation
Table of Contents
- Basic Protocol 1: Manual Procedure for 5′‐Iodination of Oligodeoxyribonucleotides on a Solid Support
- Alternate Protocol 1: Automated Procedure for 5′‐Iodination of Oligodeoxyribonucleotides on a Solid Support
- Reagents and Solutions
- Commentary
- Literature Cited
- Figures
- Tables
Materials
Basic Protocol 1: Manual Procedure for 5′‐Iodination of Oligodeoxyribonucleotides on a Solid Support
Materials
Alternate Protocol 1: Automated Procedure for 5′‐Iodination of Oligodeoxyribonucleotides on a Solid Support
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Figures
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Figure 4.19.1 Iodination of solid‐phase‐linked oligodeoxyribonucleotides. Abbreviations: B1 and B2 , thymine and/or N ‐protected nucleobases; CPG, long‐chain alkylamine controlled‐pore glass (resin). View Image -
Figure 4.19.2 RP‐HPLC profiles of 5′‐d(NGTAGGCAAGAGT) before and after 5′‐iodination. From top to bottom, N = T, C, A, and G, respectively. Reprinted from Miller and Kool () with permission from the American Chemical Society. View Image
Videos
Literature Cited
| Dimitrijevich, S.D., Verheyden, J.P.H., and Moffatt, J.G. 1979. Halo sugar nucleosides. 6. Synthesis of some 5′‐deoxy‐5′‐iodo and 4′,5′‐unsaturated purine nucleosides. J. Org. Chem. 44:400‐406. | |
| Miller, G.P. and Kool, E.T. 2002. A simple method for electrophilic functionalization of DNA. Org. Lett. 4:3599‐3601. | |
| Xu, Y. and Kool, E.T. 1997. A novel 5′‐iodonucleoside allows efficient non‐enzymatic ligation of single‐stranded and duplex DNAs. Tetrahedron Lett. 38:5595‐5598. | |
| Xu, Y. and Kool, E.T. 1998. Chemical and enzymatic properties of bridging 5′‐S‐phosphorothioester linkages in DNA. Nucl. Acids Res. 26:3159‐3164. | |
| Key References | |
| Miller and Kool, 2002. See above. | |
| Reports a method to iodinate the 5′‐carbon of oligo‐deoxyribonucleotides on a solid support. |
