Compositions and methods for covalently immobilizing an (73) Assignee: Becton Dickinson and Company, oligonucleotide onto a polymer-coated bead or similar struc Franklin Lakes, N.J. ture are described. Specifically, the polymer-coated bead or similar structure possesses a large number of activatable (21) Appl. No.: 331,296 moieties, preferably primary and secondary amines. An oligonucleotide is activated with a monofunctional or mul 22 Filed: Oct. 28, 1994 tifunctional reagent, preferably the homotrifunctional reagent cyanuric chloride. The resultant covalently immo Related U.S. Application Data bilized oligonucleotides on the beads or similar structures can serve as nucleic acid probes on solid supports, and 63 Continuation of Ser. No. 42,442, Mar. 11, 1993, abandoned, hybridization assays can be conducted wherein specific which is a continuation of Ser. No. 522,442, May 11, 1990, abandoned. target nucleic acids are detected in complex biological samples. The beads or similar structures can be employed 51 Int, Cl." ................ C12Q 1/70; C12O 1/68; free in solution, such as in a microtiter well format; in a G01N3/53; G01N3/566 flow-through format, such as in a column; or in a dipstick, 52 U.S. Cl. .................................. 536/2.2.1; 435/5; 435/6; including a multisite indicator card wherein multiple beads 435/7.1; 436/501 possessing oligonucleotides with different sequences or 58) Field of Search .................... 435/6, 5, 71; 436/501; specificities can be closely aligned whereby a multiplicity of 536/22.1 pathogens can be detected in a single biological sample. Additionally, dichlorotriazine oligonucleotides and pro (56) References Cited cesses for activating oligonucleotides by treatment with U.S. PATENT DOCUMENTS cyanuric chloride are included in the present invention.