Ligation of an oligo containing a 5'-azide with an oligo containing a 3'-propargyl group using Click Chemistry leads to a triazole linkage that has been shown to have in vivo biocompatibility. This technique has been used to synthesize DNA constructs up to 300 bases in length. When the resultant triazole linkage was placed in a PCR template, various polymerases were able to copy the sequence correctly. The linkage has also been shown to be compatible with transcription and rolling circle amplification, as well as gene expression in E. coli. In the RNA world, a hammerhead ribozyme containing the triazole linkage at the substrate cleavage site has been shown to retain its activity. A large variety of applications is envisaged for this biocompatible chemical ligation. Support for this technology is offered with the help of Tom Brown's group at the University of Southampton.
Details
Usage
Coupling: No changes needed from standard method recommended by synthesizer manufacturer.
Deprotection: Cleavage of the oligonucleotide from this support requires 2 hours at room temperature with ammonium hydroxide. Complete the deprotection as required by the nucleobases.
Specifications
Storage
Freezer storage, -10 to -30°C, dry
References
REFERENCES - CLICK LIGATION|(1) A.H. El-Sagheer, A.P. Sanzone, R. Gao, A. Tavassoli, and T. Brown, Proc Natl Acad Sci U S A, 2011, 108, 11338-43.|(2) A.H. el-Sagheer, and T. Brown, Chem Commun (Camb), 2011, 47, 12057-8.|(3) A.P. Sanzone, A.H. El-Sagheer, T. Brown, and A. Tavassoli, Nucleic Acids Res, 2012.|(4) A. Dallmann, et al., Chemistry, 2011, 17, 14714-7.|(5) A.H. El-Sagheer, and T. Brown, Proc Natl Acad Sci U S A, 2010, 107, 15329-34.
