Method and means for annealing complementary nucleic acid molecu

Chemistry: electrical and wave energy – Processes and products – Electrostatic field or electrical discharge

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204299R, 204301, 435289, 435287, 435 6, 935 78, C12N 1500

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047879637

ABSTRACT:
Method and means for accelerating the rate of hybridization of nucleic acid probes with complemental target sequences in probe assays of the filter binding or sandwich filter binding formats. The nucleic acid probes are electrophoretically concentrated at membrane means including at least a dialysis type membrane and to which the target sequences are bound. The rate of hybridization can be further enhanced by means for moving concentrated unannealed probe molecules successively in various directions along the surface of the membrane means to which the target sequences are bound. The invention further includes method and means for electrophoretically removing from the membrane means unannealed probe molecules which became adsorbed to the membrane means during hybridization. In one embodiment of the invention the membrane means comprises a cellulose dialysis membrane in laminate relation with a nitrocellulose or nylon filter having target sequence molecules bound to the surface thereof facing the dialysis membrane. In another embodiment of the invention the membrane means comprises a dialysis membrane to one side of which the target sequence molecules are bound.

REFERENCES:
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Meinkoth, J. "Hybridization of Nucleic Acids Immobilized on Solid Supports" Analytical Biochemistry 138, pp. 267-284 (1984).
Thomas, P. "Hybridization of Denatured RNA and Small DNH Fragments Transferred to Nitrocellulose", Proc. Natl. Acad. Sci. USA vol. 77, No. 9, pp. 5201-5205 (1980).
Mulcahy L., "DNA Probes: An Overview" ACPR, Nov. 1986, pp. 14-19.
Wetmur, V. "Acceleration of DNA Renaturation Rates" Biopolymers, vol. 14, pp. 2517-2524 (1975).
Chang, C. T. et al., "Effects of Microscopic and Macroscopic Viscosity on the Rate of Renaturation of DNA" Biopolymers, vol. 13, pp. 1847-1858 (1974).
Kohne, D. "Application of DNA Probe Tests to the Diagnosis of Infectious Disease" ACPR, Nov. 1986, pp. 20-29.

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