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Cell Biology Poster Session






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Tethering of Oligo DNA Probes to Polyethylene-co-acrylic Acid a Superior Support For Nucleic Acid Hybridization

Zhang, P. (Department of Chemistry and Biochemsitry, University of Southern Mississippi, USA)
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)

Contact Person: Ping Zhang (pzhang@ocean.st.usm.edu)


Abstract

Polyethylene-co-acrylic acid, PEAA, (20 wt% acid) is commercially available in pellet and film. The film has relatively few carboxylate functions on its surface, but the density of these may be increased by treatment with strong base above the glass transition temperature or by treatment with thionyl chloride and subsequent hydrolysis. DNA oligos synthesized with C12-alkyl amino tails may be exclusively end-tethered to PEAA using either conventional carbodiimide chemistry or unconventional subsurface amide formation if the acid chloride of PEAA is used. Both immobilization chemistries may be combined on the same film to yield higher oligo surface concentration. The carbodiimide reaction takes place at a pH well below the pKa of the oligo's amine tail because of rapid amine deprotonation kinetics. Radioassay results show PEAA to be virtually immune to non-specific binding of DNA. Rapid identification of SLT-II gene from E.coli via hybridization and staining with ethidium bromide was carried out on PEAA after asymmetric PCR amplification of the gene without the need for labeled probes or surface fouling agents.

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Poster Number PAzhang0791
Keywords: dna, immobilization, carbodiimide, acrylic acid, hybridization


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Zhang, P.; Fawcett, N.C.; Craven, R.D.; Evans, J.A.; (1998). Tethering of Oligo DNA Probes to Polyethylene-co-acrylic Acid a Superior Support For Nucleic Acid Hybridization. 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/zhang0791/index.html
© 1998 Author(s) Hold Copyright