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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, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99%
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
Supelco
Methanol, Pharmaceutical Secondary Standard; Certified Reference Material
Imidazole, European Pharmacopoeia (EP) Reference Standard
USP
Imidazole, United States Pharmacopeia (USP) Reference Standard
Supelco
Water, suitable for ion chromatography
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Methanol, suitable for NMR (reference standard)
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Imidazole, ReagentPlus®, 99%
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Methylamine solution, 33 wt. % in absolute ethanol ((denatured with 1% toluene))
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Triethylamine, ≥99.5%
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Benzyl azide solution, ~0.5 M in dichloromethane, ≥95.0% (HPLC)
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Imidazole, ≥99% (titration), crystalline
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Imidazole, ACS reagent, ≥99% (titration)
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Imidazole, Molecular Biology, ≥99% (titration)
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Water, for embryo transfer, sterile-filtered, BioXtra, suitable for mouse embryo cell culture
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Water, PCR Reagent
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Water, sterile-filtered, BioReagent, suitable for cell culture
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Trichloroacetic acid, suitable for electrophoresis, suitable for fixing solution (for IEF and PAGE gels), ≥99%
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Water, Nuclease-Free Water, for Molecular Biology
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Trichloroacetic acid, ACS reagent, ≥99.0%
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Triethylamine, ≥99%
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Triethylamine, ≥99.5%
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Trichloroacetic acid, BioXtra, ≥99.0%
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Trichloroacetic acid, ≥99.0% (titration)
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Azidomethyl phenyl sulfide, 95%
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Methylamine solution, 2.0 M in THF
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Imidazole, BioUltra, ≥99.5% (GC)
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Imidazole, puriss. p.a., ≥99.5% (GC)