Cell Biology Poster Session
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Hurt, T (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)
Zhang, P (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)
Evans, J.A. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA.)
Fawcett, N.C. (Department of Chemistry and Biochemistry, University of Southern Mississippi, USA)
Abstract
Many DNA hybridizations carried out on solid supports are plagued by problems of non-specific binding of target. In the past, supports had to be treated with a prehybridization solution of materials such as BSA, polyvinylpyrrolidone, DNA and other agents. Without using prehybridization mixes, we have carried out DNA hybridizations of oligonucleotide target to small complementary oligonucleotides coupled to polyethylene-co-acrylic acid (PEAA) chips. In order to test the possible non-specific binding of target to the PEAA polymer bound probe we examined its ability to bind non-complementary labelled target. The test was done with two sets of two different probes bound to separate PEAA polymer chips. The probes were a 30mer E20) and SLTC (the complement to SLT). Hybridizations were in phosphate buffer alone. We found that the non-complementary target-probe tests had close to background binding of radiolabelled probe (ie. 14 cpm above a blank) while the complementary target-probe tests had significantly higher probe bound (ie 312 cpm above a blank). This indicated that PEAA chips had little non-specific binding of oligonucleotide.
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Poster Number PApruett0787
Keywords: hybridization, oligonucleotide, support, polymer, probe
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