Cell Biology Poster Session |
Fawcett, N.C. (Department of Chemistry and Biochemsitry, University of Southern Mississippi, USA) Craven, R.D. (Department of Chemistry and Biochemsitry, University of Southern Mississippi, USA) Evans, J.A. (Department of Chemistry and Biochemsitry, University of Southern Mississippi, USA) Abstract The acid chloride of polyethylene-co-acrylic acid reacts spontaneously with synthesized oligo DNA substituted on its 5'-end with a C12-alkyl amino tail in aqueous pH 11, phosphate buffer. A primary amide bond to the DNA results exclusively via the DNA's 5'-end. The reaction proceeds spontaneously when the acid chloride film is immersed for several hours in an oligo solution. The reaction succeeds in spite of the normally rapid, exothermic hydrolysis of acid chloride. A low surface concentration of tethered oligo results, but virtually every oligo is active toward hybridization. Hybridization to the tethered oligo is easily detected by staining with ethidium bromide and observing fluorescence from the uv illuminated surface. The reaction succeeds because the acid chloride groups are located beneath the surface of the solid phase where they are protected from hydrolysis by the hydrophobic property of the polymer. The acid chloride polymer is stable, even after long periods in water. The alkyl amine tails on the oligos prefer the hydrophobic solid phase to the aqueous phase and thus penetrate the solid spontaneously where they react to form amide. Spectroscopic evidence is consistent with the reaction mode proposed.
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Zhang, P.; Fawcett, N.C.; Craven, R.D.; Evans, J.A.; (1998). A Solid Support For Tethering 5'-aminoalkyl-substituted Oligo Dna Exclusively Via Its 5' End From Aqueous Solution Without Reagents. Presented at INABIS '98 - 5th Internet World Congress on Biomedical Sciences at McMaster University, Canada, Dec 7-16th. Available at URL http://www.mcmaster.ca/inabis98/cellbio/zhang0789/index.html | ||||||||
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