Photocleavable PC Biotin Oligo Modification

Bio-Synthesis offers photocleavable PC Biotin Oligo Modification at various scales. This light activatable PC biotin modified oligonucleotide contained a photocleavable spacer between the biotin group and the DNA bases. The biotin group is separated from the 5' end nucleotide base by the photo-cleavable group and a long chain spacer arm. The photocleavable group located on the 5'-phosphate can be selectively cleaved by illumination with UV light at 300-350 nm, thereby releasing the biotin to produce a 5' phosphorylated oligo. Photo controlled oligonucleotide allow researcher to capture oligonucleotide or PCR products from a crude mixture using streptavdin mediated affinity capture and release of biotinylated oligos or PCR products in purification or for diagnostic applications. They can also be used to isolate different kinds of DNA or RNA macromolecular complexes, such as nucleosomes² and chromatin³.

These ligh controlled oligonucleotide can be use as caged oligonucleotides. Caging allows the biological activity of a species to be suppressed until released by an external agent - in this case a flash of UV light. This permits researchers exquisite spatial and temporal control of the concentration of the active species. It could be used to tether an antisense oligo to a dextran which is too large to pass through the nuclear pores. Photocleavage would then be used to release the oligo, thereby decoupling antisense effects within the cytoplasm from antisense effects within the nucleus⁴.

Contact Bio-Synthesis for Photocleavable PC Biotin Modified Oligo Service.

Purification:

Photocleavable PC biotin modified oligonucleotide can be complished by various methods. Desalting or cartridge purification is acceptable, however, additional purificaiton by HPLC is strongly recommended.

Price:

Price/modification. 10%-15% discount for more than two spacer modifications.

Cleavage Protocol

Optimal cleavage is obtained with exposure to long-wave UV light in the 300-350 nm spectral range. Cleavage releases the oligo with a 5'-phosphate group.

References:

1. Olejnik, J., Krzymanski-Olejnik, E., Rothschild, K.J. Photocleavable biotin phosphoramidite for 5’-end-labeling, affinity purification and phosphorylation of synthetic oligonucleotides. Nucleic Acids Res. (1996), 24: 361-366.
2. Shimkus, M., Levy, J., Herman, T. A chemically cleavable biotinylated nucleotide: usefulness in the recovery of protein-DNA complexes from avidin affinity columns. Proc. Natl. Acad. Sci. USA (1985), 82: 2593-2597.
3. Dawson, B.A., Herman, T., Haas, A.L., Lough, J. Affinity isolation of active murine erythroleukemia cell chromatin: uniform distribution of ubiquitinated histone H2A between active and inactive fractions.J. Cell Biochem. (1991), 46: 166-173.
4. Casey, J.P., Blidner, R.A., Monroe, W.T. Caged siRNAs for Spatiotemporal Control of Gene Silencing..Mol. Pharmaceutics (2009), 6: 669-685.