Abstract: Double hybrid or multihybrid probes and compositions are usefully combined with capture assay and immobilization to provide for detection processes in which target polynucleotides can be detected or the presence or absence of genetic mutations or defects in genetic material can be determined. The capture assay involves capturing a hybrid structure, e.g., single hybrid, double hybrid or multihybrid, or capturing a complex formed by reacting a hybrid structure with a complex forming moiety, e.g., protein, such as a binding protein including an antibody. Immobilization can also be employed prior to hybridization or complexation in which instance a polynucleotide probe can be fixed to a matrix or solid support, e.g., natural or synthetic. Capture and immobilization can be carried out using direct and indirect binding and attachment techniques. Targets can be detected directly or indirectly by using a signal generating moiety and labels.

Abstract: Disclosed is a nucleic acid hybridization assay composition for detecting the presence of absence of a target oligo- or polynucleotide in a sample. The composition comprises: a solid matrix having at least one surface which is substituted with a first intercalator capable of binding dsDNA dsRNA, or DAN-RNA hybrids; a second intercalator, which may or may not comprise at least one fluorophore, said intercalator or said fluorophore each acting as either an energy donor or an energy acceptor; and an oligo- or polynucleotide probe which is specifically hybridizable with the target oligo- or polynucleotide and has directly or indirectly bound thereto, at least one lanthanide metal chelate or at least one fluorophore, each acting as either an energy donor or an energy acceptor. Also disclosed are a method and kit for its use.

Abstract: The present invention provides a method for detecting the presence of a target polynucleotide in a sample. The method comprises the steps of (a) contacting the sample under hybridizing conditions with (i) a single-stranded polynucleotide probe capable of hybridizing to the target polynucleotide and comprising a polynucleotide and at least one intercalating molecule attached to a nucleotide of the polynucleotide by means of a linker arm, and (ii) a background-reducing reagent which chemically modifies the intercalating molecule when the probe to which it is attached is single-stranded; and (b) detecting a property change resulting from the intercalation of the intercalating molecule into a target-probe hybrid, thereby detecting the target polynucleotide. The intercalating molecule which is part of the polynucleotide probe induces a change in a property, in either the probe, the target polynucleotide or a target-probe hybrid.

Abstract: Procedures and compositions for forming and stabilizing non stable branch migrated oligo- and polydeoxynucleotides utilizing a displacer which is either single stranded or partially double stranded and hybridized to a linker strand. The displacer may contain one or more modified nucleotides.

Abstract: Compounds having the structure: ##STR1## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C.sup.1' -position of the sugar moiety, provided that when B is purine or 7-deazapurine, it is attached at the N.sup.9 -position of the purine or 7-deazapurine and when B is pyrimidine, it is attached at the N.sup.

Abstract: Compounds having the structure: ##STR1## wherein B represents 7-deazapurine, or pyrimidine moiety covalently bonded to the C.sup.1 '-position of the sugar moiety, provided that when B is 7-deazapurine, it is attached at the N.sup.9 -position of the 7-deazapurine and when B is pyrimidine, it is attached at the N.sup.1 -position;wherein A represents a moiety consisting of at least three carbon atoms which is capable of forming a detectable complex with a polypeptide when the compound is incorporated into a double-stranded ribonucleic acid, deoxyribonucleic acid duplex, or DNA-RNA hybrid;wherein the dotted line represents a chemical linkage joining B and A, provided that if B is 7-deazapurine, the linkage is attached to the 7-position of the deazapurine, and if B is pyrimidine, the linkage is attached to the 5-position of the pyrimidine; andwherein each of x, y and z represents ##STR2## either directly, or when incorporated into oligo- and polynucleotides, provide probes which are widely useful.

Abstract: Disclosed is an assay system including a compound comprising an analyte-specific moiety having substituted thereon a polymer comprising plurality of self-quenching emitter moieties and a plurality of isocharged functionality separating the emitter moieties. The present invention provides compounds that overcome the undesirable effects of self-quenching when multiple emitter moieties are used for labelling of assay reagents. Avoidance of this self-quenching phenomenon by the compounds of the invention makes it possible to introduce a more concentrated degree of labelling on to analyte-specific molecules such as oligo nucleotide probes, antibodies and other specific binding proteins and analyte-specific polysaccharides. Therefor, it is possible to effect greater assay sensitivity because the number of labels per recognition molecule(analyte-specific moiety) can be increased beyond the point previously possible without the reduction in signal caused by self-quenching.

Abstract: Compounds having the structure: ##STR1## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C.sup.1' -position of the sugar moiety, provided that when B is purine or 7-deazapurine, it is attached at the N.sup.9 -position of the purine or 7-deazapurine and when B is pyrimidine, it is attached at the N.sup.

Abstract: This invention provides methods and compositions for the detection of a target genetic material having a desired base sequence or gene. Also disclosed are methods and compositions for the detection of mutants. The methods and compositions are based in part upon techniques which utilize two labeled single-stranded polynucleotide segments which are complementary to different portions of the genetic material. The methods and compositions of this invention result in the formation of a double hybrid and/or multihybrid, and can furthermore be employed in an attached system, i.e., a matrix-bound entity capable of binding to a polynucleotide probe entity.

Abstract: The present invention provides nucleotides and polynucleotides which are chemically modified or labeled so as to be capable of ready detection when attached to and/or incorporated in nucleic acid material. More particularly, this invention provides a nucleotide selected from the group consisting of (i) a ribonucleotide having the formula, ##STR1## wherein PM is a phosphate moiety, SM is a sugar moiety, BASE is a pyrimidine, purine or 7-deazapurine moiety, and Sig is a saccharide moiety. PM is attached at the 2',3' or 5' position of SM, BASE is attached to the 1' position of SM from the N.sup.1 position when BASE is a pyrimidine or covalently attached to SM; and (ii) a nucleotide having the formula, ##STR2## wherein PM, SM, BASE and Sig are as defined above but PM is attached to the 3' or the 5' position of SM when said nucleotide is a deoxyribonucleotide and at the 2', 3' or 5' position when said nucleotide is a ribonucleotide, BASE is attached to the 1' position of SM from the N.sup.

Abstract: The present invention provides nucleotides and polynucleotides which are chemically modified or labeled so as to be capable of ready detection when attached to and/or incorporated in nucleic acid material. More particularly, this invention provides a nucleotide having the formulaPM-SM-BASE-Sigwherein PM is a phosphate moiety, SM is a sugar moiety and BASE is a pyrimidine, purine or 7-deazapurine moiety. PM is attached at the 3' or the 5' position of SM when the nucleotide is a deoxyribonucleotide and at the 2', 3' or 5' position when the nucleotide is a ribonucleotide. BASE is attached to the 1' position of SM from the N.sup.1 position when BASE is a pyrimidine or the N.sup.9 position when BASE is a purine or a 7-deazapurine. Sig is a detectable moiety that is covalently attached to BASE at a position other than the C.sup.5 position when BASE is a pyrimidine, at a position other than the C.sup.8 position when BASE is a purine and at a position other than the C.sup.7 position when BASE is a 7-deazapurine.

Abstract: A detectable molecule of the formulaA.sup.3 --(--X--R.sup.1 --E--Det.sup.b).sub.mwhere A.sup.3 is A.sup.2 or a polymer, where A.sup.3 has at least one modifiable reactive group selected from the group consisting of amino, hydroxy, cis OH, halides, aryl, imidazoyl, carbonyl, carboxy, thiol or a residue comprising an activated carbon; --X-- is selected from the group consisting of ##STR1## or a C.sub.1 -C.sub.10 branched or unbranched alkyl or aralkyl, which may be substituted by --OH; --Y-- is a direct bond to --E--, or --Y-- is --E--R.sup.2 -- where R.sup.2 is a C.sub.1 -C.sub.10 branched or unbranched alkyl; Z.sub.a is chlorine, bromine or iodine; E is O, NH or an acyclic divalent sulfur atom; Det.sup.b is a chemical moiety capable of being detected, preferably comprising biotin or a metal chelator of the formula: ##STR2## or the 4-hydroxy or acyloxy derivative thereof, where R.sup.3 is C.sub.1 -C.sub.4 alkyl or CH.sub.

Abstract: An oligo- or polynucleotide having at least one biotin directly or indirectly attached to each of the 5' and 3' end nucleotides thereof and a nucleic acid hybridization assay system for use therewith. The biotins are optionally attached through linkage groups that do not interfere with hybridization. The biotins can further serve to attach a biotinylated polymer.

Abstract: A time-resolved fluorometer, having a light tight enclosure, for detecting the presence of an analyte in a sample. Within the light tight enclosure are a pulsed dye laser that produces a pulsed light beam for sample excitation, a light tight sample excitation station through which samples, treated with a reagent composition, are passed into the path of the pulsed light excitation beam to produce a delayed fluorescence emission, a fused silica lens system through which the delayed fluorescence emission passes and an assembly which selectively amplifies, counts and characterizes the resulting emissions. The operation and coordination of the time resolved fluorometer are under computerized control as are the readings reported. Significant improvements relate to the fused silica lens systems and interference filters, cooling of the emission measurement apparatus and particularly the improved performance resulting from the combination of these aspects.

Abstract: Disclosed is an improvement relating to adhesion of samples onto a surface for nucleic acid hybridization assay to detect a target polynucleotide. In one aspect, the invention provides a method of adhering a tissue, cell or other target polynucleotide-containing sample to a substrate under nucleic acid hybridization assay compatible conditions. In another aspect, the method comprises a hybridization assay procedure. Also disclosed are kits for performance of such procedures.

Abstract: In vivo labelled polynucleotides, processes for in vivo labelling of polynucleotides, and detection methods and kits characterized by those labelled polynucleotides. The in vivo on biologically-labelled polynucleotides of this invention are useful in the detection of various analytes and in other laboratory, industrial and medical applications.