| Reference | [1]. Esters, N.H.S.,<br />
The copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction between azides and alkynes to form 1,2,3-triazoles, as reported1 by Sharpless, was found to be so exquisitely regioselective and efficient at even the most mild conditions that Sharpless coined the term ‘Click Chemistry’ to describe it. The use of this method for DNA modification has been somewhat delayed by the fact that copper ions damage DNA, typically yielding strand breaks.2 As these problems have now been overcome by the use of copper(I)-stabilizing ligands (e.g., tris(benzyltriazolylmethyl)amine, TBTA3), Carell et al. and Seela et al. discovered that the CuAAC reaction can be used to functionalize alkyne-modified DNA nucleobases with extremely high efficiency.4 At Glen Research, our goal was to offer a copper-free click phosphoramidite reagent with the following properties: Simple to use Stable in solution on the synthesizer Stable to ammonium hydroxide and AMA< Excellent click performance in 17 hours or less at room temperature From the variety of cyclooctyne-based copper-free click reagents so far described, we have chosen to offer compounds based on a dibenzo-cyclooctyne (DBCO) structure.<br />
Click and Copper-free Click Chemistry Monomers, Supports, and Kits.<br />
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[2]. CeP, A.P., 1999. from Berry & Associates. Bioconj. Chem, 10, pp.261-270.<br />
Abstract: Given the large and growing body of work on anthraquinone-modified oligonucleotides, it is surprising that there are no commercially available phosphoramidites for their incorporation. We have chosen to offer a suite of new anthraquinone-bearing phosphoramidites.<br />
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[3]. De Clercq, E., Descamps, J., Barr, P.J., Jones, A.S., Serafinowski, P., Walker, R.T., Huang, G.F., Torrence, P.F., Schmidt, C.L., Mertes, M.P. and Kulikowski, T., 1979.<br />
Comparative study of the potency and selectivity of anti-herpes compounds.<br />
ABSTRACT: Several anti-herpes compounds, including phosphonoacetic acid, 9-(2-hydroxyethoxymethyl) guanine, 9-β-D-arabinofuranosyladenine, 1-β-D-arabinofuranosylthymine, 5-iodo-5′-amino-2;5′-dideoxyuridine, 5-iodo-2′-deoxyuridine, 5-bromo-2′-deoxycytidine and various other 5-substituted 2′-deoxyuridines and 2′-deoxycytidines were compared in vitro (primary rabbit kidney cells) for both antiviral activity (based on the ID50 required to inhibit the cytopathogenicity of the KOS strain of type 1 herpes simplex virus) and antimetabolic activity (based on the ID50 required to inhibit the incorporation of 2′-deoxyuridine into host cell DNA). Of the whole set of compounds tested, E-5-(2-bromovinyl)- and E-5-(2-iodovinyl)-2′-deoxyuridine emerged as both the most potent and the most selective anti-herpes agents.<br />
In Antimetabolites in biochemistry, biology and medicine (pp. 275-285). Pergamon.<br />
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[4]. Hall, L.M., Gerowska, M. and Brown, T., 2012.<br />
A highly fluorescent DNA toolkit: synthesis and properties of oligonucleotides containing new Cy3, Cy5 and Cy3B monomers.<br />
Abstract: Cy3B is an extremely bright and stable fluorescent dye, which is only available for coupling to nucleic acids post-synthetically. This severely limits its use in the fields of genomics, biology and nanotechnology. We have optimized the synthesis of Cy3B, and for the first time produced a diverse range of Cy3B monomers for use in solid-phase oligonucleotide synthesis. This molecular toolkit includes phosphoramidite monomers with Cy3B linked to deoxyribose, to the 5-position of thymine, and to a hexynyl linker, in addition to an oligonucleotide synthesis resin in which Cy3B is linked to deoxyribose. These monomers have been used to incorporate single and multiple Cy3B units into oligonucleotides internally and at both termini. Cy3B Taqman probes, Scorpions and HyBeacons have been synthesized and used successfully in mutation detection, and a dual Cy3B Molecular Beacon was synthesized and found to be superior to the corresponding Cy3B/DABCYL Beacon. Attachment of Cy3, Cy3B and Cy5 to the 5-position of thymidine by an ethynyl linker enabled the synthesis of an oligonucleotide FRET system. The rigid linker between the dye and nucleobase minimizes dye–dye and dye–DNA interactions and reduces fluorescence quenching. These reagents open up new future applications of Cy3B, including more sensitive single-molecule and cell-imaging studies.<br />
Nucleic acids research, 40(14), pp.e108-e108.<br />
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[5]. Ren, X., El-Sagheer, A.H. and Brown, T., 2015.<br />
Azide and trans-cyclooctene dUTPs: incorporation into DNA probes and fluorescent click-labelling.<br />
Abstract: 5-Azidomethyl dUTP and two 5-trans-cyclooctene dUTPs with different linkers between the TCO and the uracil base have been incorporated into DNA by primer extension, reverse-transcription and PCR amplification. For azidomethyl dUTP the PCR reaction was successful even when the modified dUTP was not supplemented with dTTP. In one case 335 azidomethyl dU residues were incorporated into the 523 base pair amplicon using this methodology. 5-Azidomethyl dUTP was found to be a better substrate for DNA polymerases than the trans-cyclooctene dUTPs. However, the inverse electron demand Diels–Alder reaction between cyclooctene DNA and a tetrazine Cy3-dye was more efficient than the strain-promoted reaction between azide DNA and a bicyclo [6.1.0] non-4-yne Cy3 dye.<br />
Analyst, 140(8), pp.2671-2678.
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