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  • High-throughput chemical modification of oligonucleotides for systematic structure-activity relationship evaluation.

High-throughput chemical modification of oligonucleotides for systematic structure-activity relationship evaluation.

Bioconjugate chemistry (2014-11-15)
Daniel Zewge, Francis Gosselin, Denise M Kenski, Jenny Li, Vasant Jadhav, Yu Yuan, Sandhya S Nerurkar, David M Tellers, W Michael Flanagan, Ian W Davies
ABSTRACT

Chemical modification of siRNA is achieved in a high-throughput manner (96-well plate format) by copper catalyzed azide-alkyne cycloadditions. This transformation can be performed in one synthetic operation at up to four positions with complete specificity, good yield, and acceptable purity. As demonstrated here, this approach extends the current synthetic options for oligonucleotide modifications and simultaneously facilitates the systematic, rapid biological evaluation of modified siRNA.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Imidazole, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
Triethylamine, BioUltra, ≥99.5% (GC)
Pure Water Density Standard, UKAS ISO/IEC17025 and ISO Guide 34 certified, density: 0.9982 g/mL at 20 °C, density: 0.9970 g/mL at 25 °C
Pure Water Density Standard, UKAS ISO/IEC17025 and ISO Guide 34 certified, density: 0.9982 g/mL at 20 °C, density: 0.9970 g/mL at 25 °C
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Imidazole, Vetec, reagent grade, 98%
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Trichloroacetic acid, BioXtra, ≥99.0%
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Trichloroacetic acid, ≥99.0% (titration)
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Methylamine solution, 2.0 M in methanol
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2,6-Lutidine, analytical standard
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Imidazole, BioUltra, Molecular Biology, ≥99.5% (GC)
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Water, tested according to Ph. Eur.
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Methanol, anhydrous, 99.8%
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2,6-Lutidine, ReagentPlus®, 98%
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Methylamine solution, 33 wt. % in absolute ethanol ((denatured with 1% toluene))
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Methanol, purification grade, 99.8%
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Methanol, ACS reagent, ≥99.8%
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Methanol, ACS spectrophotometric grade, ≥99.9%
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Methanol, suitable for HPLC, ≥99.9%
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Water, for embryo transfer, sterile-filtered, BioXtra, suitable for mouse embryo cell culture
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Imidazole, ReagentPlus®, 99%, Redi-Dri, free-flowing
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2,6-Lutidine
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Methylamine solution, 40 wt. % in H2O
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Triethylamine, ≥99.5%
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Triethylamine, ≥99%