Abstract: Polynucleotides containing abasic, cleavable sites are provided. These polynucleotides are useful in a variety of biochemical and chemical contexts, particularly in solid phase nucleic acid hybridization assays because a captured probe can be released from the support. The polynucleotides have the structure ##STR1## where R is selected from the group consisting of 2-nitrobenzyl, 4-penten-1-yl, ##STR2## where R', R.sub.i and R.sub.j are as defined herein.

Abstract: Hydroxyl-protecting groups orthogonally removable by reduction with a liquid reducing agent are disclosed. The novel hydroxyl-protecting groups are particularly useful in the chemical synthesis of linear and branched oligonucleotide structures, as they are readily removed from the protected molecule with mild reagents such as dithionite. Examples of such hydroxyl-protecting groups include the 2-methylene-9,10-anthraquinone (Maq) carbonate ester and the p-nitrobenzyl carbonate ester.

Abstract: Novel DNA probe sequences for detection of CMV in a sample in a solution phase sandwich hybridization assay are described. Amplified nucleic acid hybridization assays using the probes are exemplified.

Abstract: Novel methods for assaying a nucleic acid analyte are provided, which employ polynucleotides having oligonucleotide sequences substantially homologous to a sequence of interest in the analyte, where the presence or absence of hybridization at a predetermined stringency provides for the release of a label from a support. Particularly, various techniques are employed for binding a label to a support, whereupon cleavage of either a single or double strand, a label may be released from a support, where the release of the label can be detected as indicative of the presence of a particular oligonucleotide sequence in a sample. The method finds use in diagnosis of disease, genetic monitoring, and analysis of nucleic acid mixtures.

Abstract: A modified polynucleotide containing at least one cleavable or abasic site as shown below. ##STR1## DNA.sub.1 is a first segment of DNA; DNA.sub.2 is a second segment of DNA; and R.sub.m is C.sub.1 to C.sub.16 alkylene or an oxytheylene oligomer --(CH.sub.2 CH.sub.2 O).sub.z -- where z is an interger in the range of 1 to 16 inclusive, and R.sub.n is selected from the group consisting of ##STR2## Such polynucleotides are useful in solid phase hybridizations because they permit the release of a label from the solid support after the hybridization reaction.

Abstract: Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Abstract: A self-contained assembly for assaying an analyte in a liquid sample. The pair of disc-like rotatable plates forming the assembly are relatively rotatable to align reagent reservoirs in one plate with a reaction well in the other plate, for sequential addition of multiple reagents, either in liquid or solid form, to the reaction well. In one embodiment, the reaction well includes solid-phase particles which can be transferred from the reaction well to spaced wells in the assembly by a combination of relative movement of the plate, and rotation of the entire assembly.

Abstract: Novel photolabile photochemical reagents are disclosed. The reagents are useful in a variety of biochemical and chemical contexts, including nucleic hybridization assays and chemical phosphorylation of hydroxyl-containing compounds. The reagents are particularly useful for introducing cleavable sites into oligonucleotide or polynucleotide chains, i.e., sites which are cleavable upon photolysis. The reagents are also useful in both 5'- and 3'-phosphorylation of oligonucleotide or polynucleotide chains.

Abstract: Oligomers and polymers are prepared substantially free of error sequences by sequentially adding monomers, which are terminally blocked and have active functionalities protected, to a growing chain bound to a support through a selectively cleavable linkage. After each addition, unblocked terminal groups are capped. At the completion of monomer addition, enzymatic hydrolysis interfering protecting groups are removed along with the capping group and failure sequences enzymatically degraded. The terminal blocking group may then be removed. The completed oligomer or polymer may be cleaved from the support prior or subsequent to enzymatic degradation but after completion of the sequence.

Abstract: A method of phosphorylating a nucleoside or an oligonucleotide chain having a free 2', 3' or 5' hydroxyl moiety is provided. The method involves the use of a phosphorylating reagent which is selected such that the extent of phosphorylation can be monitored colorimetrically, easily and accurately. The phosphorylating reagent contains an aromatic species such as a dimethoxytrityl group that is cleavable with acid and colorimetrically detectable upon release.

Abstract: A stable 1,2-dioxetane of the formula: ##STR1## wherein X' is a phenyl group with 0 to 2 Cl or I atoms. This 1,2-dioxetane is a double-trigger substrate. The 2-methyl-4-hydroxy-naphthyl group is first removed by horseradish peroxidase. The exposed phosphate group is then removed with alkaline phosphatase. The exposed phenoxy 1,2-dioxetane decomposes to generate chemiluminescence output in immunoassays or nucleic acid hybridization assays.

Abstract: Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Abstract: Novel methods for assaying a nucleic acid analyte are provided, which employ polynucleotides having oligonucleotide sequences substantially homologous to a sequence of interest in the analyte, where the presence or absence of hybridization at a predetermined stringency provides for the release of a label from a support. Particularly, various techniques are employed for binding a label to a support, whereupon cleavage of either a single or double strand, a label may be released from a support, where the release of the label can be detected as indicative of the presence of a particular oligonucleotide sequence in a sample. The method finds use in diagnosis of disease, genetic monitoring, and analysis of nucleic acid mixtures.

Abstract: Modified nucleotides are provided which have the structure ##STR1## wherein R.sup.1 is a reactive group derivatizable with a detectable label, R.sup.2 is an optional linking moiety including an amide, thioether or disulfide linkage or a combination thereof, R.sup.3 is hydrogen, methyl, bromine, fluorine or iodine, R.sup.4 is hydrogen, an acid-sensitive, base-stable blocking group of an acyl capping group, R.sup.5 is hydrogen or a phosphorus derivative, R.sup.6 is H, OH, or OR where R is a protecting group and x is an integer in the range of 1 and 8 inclusive. Methods of synthesizing the derivatizable nucleotide are disclosed, as are labeled polynucleotide probes prepared therefrom.

Abstract: Modified nucleotides are provided which have the structure ##STR1## wherein R.sup.1 is a reactive group derivatizabale with a detectable label, R.sup.2 is an optional linking moiety including an amide, thioether or disulfide linkage or a combination thereof. R.sup.3 is hydrogen, methyl, bromine, fluorine or iodine, R.sup.4 is hydrogen, an acid-sensitive, base-stable blocking group or an acyl capping group, R.sup.5 is hydrogen or a phosphorus derivative, R.sup.6 is H, OH, or OR where R is a protecting group and x is an integer in the range of 1 and 8 inclusive. Methods of synthesizing the derivatizable nucleotide are disclosed, as are labeled polynucleotide probes prepared therefrom.