Abstract: Enzymatic synthesis of oligonucleotides is performed by the steps of: (a) combining a primer and a blocked nucleotide in the presence of a chain extending enzyme to form a primer-blocked nucleotide product containing the blocked nucleotide coupled to the primer at its 3'-end; (b) removing the blocking group from the 3' end of the primer-blocked nucleotide product; and (c) repeating the cycle of steps (a) and (b), using the primer-nucleotide product of step (b) as the primer for step (a) in the next cycle, for sufficient cycles to form the oligonucleotide product. Cycles may optionally include the step of converting any unreacted blocked nucleotide to an unreactive form which is substantially less active as a substrate for the chain extending enzyme. Cycles may also include the step of removing the blocking group from unreacted blocked nucleotide. This step is unnecessary, however, when the same nucleotide is added in two or more successive cycles.

Abstract: The amino acid sequence of the full length version of the human neurotrophic factor receptor TRK B and a nucleotide sequence encoding it are disclosed. Transformed host cells and transgenic animals containing nucleic acid molecules encoding full length human TRK-B are disclosed. Probes, primers and antisense molecules identical and complementary to the nucleotide sequence that encodes full length human TRK-B, particularly the intracellular portion of the receptor are disclosed. Monoclonal antibodies that bind to the full length human TRK-B protein are disclosed. Methods of using and kits which include probes, primers, antisense molecules and monoclonal antibodies are disclosed.

Abstract: A waveguide binding assay method involves detecting the scattering of light directed into the waveguide, the scattering being the result of scattering labels specifically bound to the waveguide within the penetration depth of an evanescent wave. The waveguide may be transparent plastic or glass and the binding is typically by oligonucleotide hybridization or immunological capture. Light scattering labels include colloidal metals or non-metals, including gold, selenium and latex. A light absorbing member consisting of dye or concentrated particles may also be employed to enhance signal. Real-time binding and dissociation can be monitored visually or by video imaging, such as with a CCD camera and frame grabber software. Hybridization mismatches of as few as one base can be distinguished by real-time melting curves.

Abstract: The present invention provides a process for determining genotypes in highly polymorphic systems by polymerase chain reaction amplification of cDNA or genomic DNA and direct sequencing polymerase chain reaction products using oligonucleotide primers. More specifically, Class I HLA genotypes can be unambiguously determined in any subject in 16-24 hours by direct sequencing of HLA-A, HLA-B, and HLA-C transcripts enzymatically amplified and sequenced using a limited number of selected oligonucleotide.

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 oligonucleotide probes are provided having the structure ##STR1## wherein: R.sup.2 is lower alkyl; R.sup.3 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --O--, --S-- and --NR.sup.12 -- wherein R.sup.12 is hydrogen or lower alkyl; R.sup.5 is hydrogen or lower alkyl; R.sup.6 is selected from the group consisting of hydrogen, methyl, bromo and iodo; R.sup.7 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --O--, --S-- and --NR.sup.12 --, and bound to R.sup.8 through an --O--, --S-- or --NR.sup.12 -- moiety; and R.sup.8 is a protecting group that can be removed and replaced, without affecting the remainder of the compound, by reduction with a reducing agent. Methods for using the probes are provided as well.

Abstract: A nucleic acid molecule which codes for a tumor rejection antigen precursor which is in turn processed to an antigen presented by HLA-B44 molecule is described. The antigen is also described. These materials are useful in diagnostic and therapeutic methodologies. The tumor rejection antigen precursor is not tyrosinase, which has previously been identified as a tumor rejection antigen precursor processed to an antigen presented by HLA-B44.

Abstract: The invention provides a diagnostic method of determining Kell genotype by the identification of the molecular basis of a Kell polymorphism. Specifically, the invention provides a method for determining K1/K2 genotype with great accuracy, overcoming problems associated with traditional serological typing methods. The diagnostic method of the invention preferably employs amplification of K1/K2 nucleic acid sequences, and optionally employs differential cleavage of K1- and K2-specific nucleic acid sequences by a restriction enzyme. Also provided are nucleic acid oligomers useful as probes or primers for the method of the invention. Furthermore, diagnostic kits for the determination of Kell genotype are provided.

Abstract: The present invention concerns in situ polymerase chain reaction and provides methods and reagents for identifying cells containing at least one selected nucleic acid sequence which may be derived from the human immunodeficiency virus.

Abstract: Antibodies which are specific to a thermostable DNA polymerase can be used to reduce or eliminate the formation of non-specific products in polymerase chain reaction methods. These antibodies and other temperature sensitive inhibitors are effective to inhibit DNA polymerase enzymatic activity at a certain temperature T.sub.1 which is generally below about 85.degree. C. The inhibitors are irreversibly inactivated at temperature T.sub.2 which is generally above about 40.degree. C. T.sub.2 is also greater than T.sub.1. Such inhibitors can be supplied individually or in admixture with the DNA polymerase in a diagnostic test kit suitable for PCR.

Abstract: The present invention contemplates methods of decontaminating human fluids prior to processing in the clinical laboratory. The techniques handle large volumes of human serum without impairing the testing results. Novel compounds for photodecontaminating biological material are also contemplated which are compatible with clinical testing, in that they do not interfere with serum analytes.

Abstract: Oligonucleotides primers can be used to amplify and detect Hepatitis C virus nucleic acids in a process that involves reverse transcription of the viral genomic RNA to create cDNA and the subsequent amplification of the cDNA by the polymerase chain reaction. Oligonucleotide probes can be used to detect the presence of amplified DNA by hybridization. The invention provides improved methods, compositions, and kits for amplifying and detecting U.S., Japan, and HCV-C prototype nucleic acids.

Abstract: The present invention pertains to a process which can be fully automated for accurately determining the alleles of STR genetic markers. More specifically, the present invention is related to performing PCR amplification on a location of DNA, labelling the PCR products, converting the labels into a signal, removing a reproducible PCR stutter pattern from the signal by means of a computational device, and then determining the genotype of the location of the DNA. The invention also pertains to genetics applications and systems which can effectively use this genotyping information.

Abstract: Novel N-4 modified pyrimidine analogs are provided having the structure (I) or (II) ##STR1## wherein: R.sup.1 is selected from the group consisting of hydrogen, acid-sensitive, base-stable blocking groups and acyl capping groups; R.sup.2 is lower alkyl; R.sup.3 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --0--, --S-- and --NH--; R.sup.4 is ##STR2## in which R.sup.9 is hydrogen or an optionally substituted aliphatic group; R.sup.10 and R.sup.11 are hydrocarbyl or together form a mono- or polyheterocyclic ring; R.sup.5 is hydrogen or lower alkyl; R.sup.6 is selected from the group consisting of hydrogen, methyl, bromo and iodo; R.sup.7 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --O--, --S-- and --NR.sup.12 -- wherein R.sup.12 is hydrogen or lower alkyl; and R.sup.8 is a protecting group that can be removed and replaced by reduction.

Abstract: A procedure is provided for the sex determination of embryos in mammals, that is particularly exemplified on bovines. The procedure overcomes the limitations given in former techniques. It includes a new stage of polymerase chain reaction PCR method, using specific oligonucleotides in order to amplify fragments corresponding to the sampled cells and not from false ones, spuriously produced. In this way, the necessary sensitivity is obtained. The oligonucleotides sequences are particularly selected for the procedure. The procedure is used for the sex determination of embryos in mammals, and the procedure is sensitive enough to be carried out a sample containing few cells, is highly secure, and which give positive results for both female and male sex; specially applied to bovine embryos.

Abstract: The invention relates to a process for reducing RNA concentration in a mixture of biological material, comprising the steps of: (a) providing a mixture of biological material having a first concentration of RNA; (b) filtering the mixture through a diatomaceous earth material to produce a filtrate having a second concentration of RNA, wherein the second concentration is less than the first concentration; and (c) collecting the filtrate having a reduced RNA concentration.

Abstract: The present invention concerns a process for introducing non-radioactively labelled deoxynucleotides into nucleic acids and RNA molecules which at their 3' end contain at least one deoxynucleotide which carries a non-radioactive marker group.

Abstract: The synthesis and use of polymeric carriers is described which are loaded with nucleic acid building blocks which in turn contain labelling groups or precursors thereof. The polymeric carrier loaded in this way serves as a solid or liquid phase for the assembly of oligonucleotides which can be used as primers for template-dependent enzymatic nucleic acid syntheses such as for example in sequencing analysis according to Sanger and co-workers or in the polymerase chain reaction (PCR).

Abstract: Compositions and methods for detecting the conversion to mucoidy in Pseudomonas aeruginosa are disclosed. Mucoidy is a critical P. aeruginosa virulence factor in cystic fibrosis that has been associated with biofilm develoment and resistance to phagocytosis. The present invention provides for detecting the switch from nonmucoid to mucoid state as caused by the interaction of the algU gene product, algU, with RNA polymerase. Inactivation of algU results in a loss of expression of genes, such as algD, dependent on algU for transcription. Also disclosed is a novel alginate biosynthesis heterologous expression system for use in screening candidate substances that inhibit conversion to mucoidy by inhibiting the interaction of algU with the RNA polymerase holoenzyme.

Abstract: A method for determining the genotype of a gypsy moth is disclosed. This method begins with the steps of isolating genomic DNA from a candidate gypsy moth, exposing that DNA to an oligonucleotide primer selected from the group consisting of SEQ ID NOs:3, 4, 5, 6, 7 or the complement thereof and sequences sufficiently similar to SEQ ID NOs:4 and 5 or the complement thereof under conditions permitting amplification of the genomic DNA, and comparing the amplified DNA with amplified DNA produced by control samples. A genotype identity is assigned to the candidate moth according to which control sample has amplified fragments similar to those in the candidate sample. A method for obtaining DNA primers for determining the genotype of candidate moths is also disclosed.