Abstract: The present invention discloses polymerase chain reaction (PCR) based assays for the presence of the heartworm parasite Dirofilaria immitis prevalent in dogs and cats, as well as in mosquitoes, which are the transmission vector. The assays are extremely sensitive, and can be used effectively to screen large numbers of samples. The invention provides primers and kits for use in the PCR assay of the invention which are oligonucleotides or oligonucleotide derivatives specific for D. immitis. The invention further discloses methods of detecting the presence of D. immitis in a sample based on PCR which employs the primers of the invention. The invention also discloses methods for rapidly obtaining parasite DNA from blood samples and mosquitoes. The samples that may be assayed include blood samples from mammals (especially dogs and/or cats) suspected of harboring D. immitis and mosquitoes suspected of transmitting the pathogen.

Abstract: Described herein are methods for extracting DNA from serum or plasma, comprising contacting serum or plasma with alkali to yield alkalinized serum or plasma, heating the alkalinized serum or plasma to a temperature ranging from about 100 to about 110° C. for a time ranging from about 5 to about 20 minutes, centrifuging the heated alkalinized serum or plasma to yield a DNA-containing supernatant, allowing the heated alkalinized serum or plasma to cool to room temperature, or about 25° C., and recovering the DNA-containing supernatant. Also disclosed are methods for detecting a DNA-containing microorganism in serum or plasma.

Abstract: The present invention refers to a method that allows the use of filamentous bacteriophages to detect the presence of molecules of interest in biological samples. It consists of a series of combined operations, which are drawn up and defined according to the characteristics of the phages employed. These have, exposed on the surfaces of the capsid, at least one molecule, generally but not exclusively of proteic nature, capable of binding at least one molecule of interest present in the biological sample. Detection of the presence of the molecule takes place, according to the method of the invention, using the association between the molecule exposed on the surfaces of the phage and the genome of the phage itself. The ability to form specific complexes typical of the molecule exposed on the capsid enables formation of phage-molecule of interest complexes, which can be detected by means of amplification of known sequences in the phage DNA.

Abstract: The invention relates to a method for marking solid, liquid or gaseous substances, whereby the substance to be marked is provided with at least one synthetically produced nucleic acid sequence. Said nucleic acid sequence contains a first sequence section constructed with the 5′ terminal end, a second sequence comprised of at least two bases and connected to said nucleic acid sequence, and a third sequence section constructed with the 3′ terminal end and connected to the said nucleic acid sequence. In order to simplify the identification of the marking, the invention provides that a first primer group is used with a first primer section corresponding to the first sequence section and a second primer group is used with a third primer section corresponding to the third sequence section.

Abstract: A method and apparatus for detection of multiple target nucleic acids and/or antigens such as hormones, antibodies, or nerve agents in a sample, involves presenting the sample to a plurality of reporter binding sites wherein each reporter binding site comprises two partially hybridized molecules. A first of the two hybridized molecules is bound to the binding site and is complementary to a target nucleic acid or antigen, and it will therefore hybridize to the target nucleic acid or antigen and cause the release of the second hybridized molecule into the sample. The second hybridized molecule comprises a reporter nucleic acid sequence, which uniquely identifies the target nucleic acid or antigen. Subsequent PCR amplification of the unique reporter nucleic acid sequence using labeled primers results in multiple labeled copies of the unique nucleic acid sequence.

Abstract: The present invention relate to methods and compositions for simultaneously analyzing multiple different polynucleotides of a polynucleotide composition comprising multiple diverse polynucleotide sequences. The subject methods and compositions may also be applied to analyze or identify single polynucleotides; however, the subject methods and compositions are particularly useful for analyzing large diverse populations of polynucleotides, e.g., cDNA libraries. Most embodiments of the invention involve hybridizing terminus probes (of known base sequence) to adapter-modified restriction fragment generated from polynucleotide for analysis, and subsequently joining the terminus probes and internal fragment probes to each other. The terminus probe hybridizes to bases of restriction endonuclease recognition site present at the terminus of a restriction fragment generated from the polynucleotide for analysis.

Abstract: Disclosed are compositions and an in vitro method for cloning and/or amplification of nucleic acid sequences of interest. The method is based on strand displacement replication of the nucleic acid sequences by multiple priming on artificial long terminal repeat (ALTR) sequences appended to the ends of the nucleic acid molecule of interest. The nucleic acid molecules for cloning and amplification can be very long, up to 40 to 80 Kb or longer. In a preferred form of the method, a single primer is used to prime strand displacement replication at multiple sites in artificial long terminal repeat sequences, flanking a target nucleic acid, containing multiple tandem repeats of a primer complement sequence. Amplification proceeds by replication initiated at each primer and continuing through the target nucleic acid sequence. This nested replication of multiple copies significantly increases the amplification yield for extremely long nucleic acid molecules.

Abstract: The present invention utilizes the singularity of the 18S rRNA gene sequence of the Cordyceps sinensis between the NS3/NS6 primer pair as the index for distinguishing the Cordyceps sinensis from other Cordyceps species.

Abstract: The invention concerns a method for the amplification of nucleic acids using a reagent which prevents reactivation of degradation enzymes. This enables a simpler prevention of contaminations.

Abstract: Isolated nucleic acid molecules encoding Thyroid Receptor-Associated Proteins (TRAPS) are provided. TRAPS are members of protein complexes that bind to nuclear hormone receptors in a ligand-dependent manner so that the receptor, upon activation by a corresponding hormone, regulates the transcription of a particular gene. Also provided are methods of replicating and expressing such isolated nucleic acid molecules, pharmaceutical compositions comprising TRAPS, and methods of modulating gene expression via administration of therapeutically effective amounts of such pharmaceutical compositions.

Abstract: A labeled primer for use in detection of nucleic acid is described, which primer is labeled at the two ends of the oligonucleotide strand with a reporter dye molecule and a quencher molecule, and in which labeled primer, at least one base at the 3′ end is deliberately not complementary to the nucleic acid sequence to be amplified. A process for detecting a nucleic acid sequence using a labeled primer is also described. The process can also be used in amplification processes.

Abstract: Described herein is a method for selectively inhibiting the amplification of a specific DNA template during a polymerase chain reaction (PCR). In particular, the method is useful when the sequences of the desired and undesired DNA templates are similar. A set of universal primers binds to both the desired and undesired DNA templates during a PCR, resulting in the amplification of their DNA sequences. The method targets the undesired DNA template with three sets of oligonucleotide primers, one set of which is terminally modified to both prevent primer extension and increase the primer-template binding affinity. The result of these terminal modifications is the specific inhibition of the PCR amplification of the undesired DNA template, allowing the preferential amplification of the desired DNA templates.

Abstract: The present invention provides a method for amplifying a specific target nucleic acid sequence. The method comprises (1) forming a reaction mixture comprising: (i) the target sequence; (ii) primers comprising a first primer at least a portion of which at the 3′ end thereof is substantially complementary to a first segment at a first end of the target sequence, a second primer at least a portion of which at the 5′ end thereof is substantially complementary to a second segment at a second end of the target sequence, the 5′ end of the second primer being adjacent the 3′ end of the first primer, and a third primer, the third primer being substantially complementary to a segment of the second primer at the 3′ end thereof; (iii) at least four different nucleotide bases; (iv) thermostable polymerase and thermostable ligase; and (2) thermocycling the reaction mixture.

Abstract: The present invention relate to methods and compositions for simultaneously analyzing multiple different polynucleotides of a polynucleotide composition comprising multiple diverse polynucleotide sequences. The subject methods and compositions may also be applied to analyze or identify single polynucleotides; however, the subject methods and compositions are particularly useful for analyzing large diverse populations of polynucleotides, e.g., cDNA libraries. Most embodiments of the invention involve hybridizing terminus probes (of known base sequence) and internal fragment probes (of known base sequence) at adjacent positions on an adapter-modified restriction fragment generated from polynucleotide for analysis, and subsequently joining the terminus probes and internal fragment probes to each other. The terminus probe hybridizes to bases of restriction endonuclease recognition site present at the terminus of a restriction fragment generated from the polynucleotide for analysis.

Abstract: An extended fibre point (1) is coated with photbiotin and irradiated with U.V. light, causing the photobiotin to bind with the fibre pint. The fibre point is then brounght into contact with streptavidin and submerged in a solution containing dyed copies of the DNA molecule to be sequenced. Biotin is also bonded to the 5′ end of the dyed DNA molecules (4A). the biotin coupled to individual DNA molecules binds with the streptavidin at the fibre point. Light is coupled into the fibre to detect the thus obtained bonds. The evanescent field of said light excites a dyed DNA molecule to emit fluorescent light as soon as it binds with the fibre point. When a bond is deteccted, the fibre is immediately remvoded from the solution, and the fibre point is inserted into a microcapillary (5). The microcapillary is filled with a buffer solution containing exonucleases which decompose the DNA colecule base after base.

Abstract: The inventive method for assaying DNA fragments in mixture comprises step 1 of ligating different oligomers hybridizable to primers of the same melting temperature and the same length to individual groups of DNA fragments in a set of DNA fragments; step 2 of mixing together the groups of DNA fragments ligated with the oligomers; step 3 of simultaneous PCR of the groups of DNA fragments ligated with the oligomers in one receptacle by using the primers being complementary to the oligomers and corresponding to the individual groups; and step 4 of detecting PCR amplified DNA fragments; characterized in that the method enables the comparison of plural samples under no influence of PCR reproducibility.

Abstract: An open system is provided for performing a submicroliter reaction. An open system can contain a solid support having a target site for performing the reaction; a liquid dispensing system such as a nanoliter dispensing pipette for dispensing a submicroliter amount of a liquid to the target site; a temperature controlling device for regulating the temperature of the support; and means for controlling the amount of liquid dispensed, which corresponds to the amount of liquid that evaporates from the target site. Also provided is an open system, including a solid support having a target site; a liquid dispensing system, which can dispense a liquid to the target site; a temperature controlling system, which regulates the temperature of the solid support; and an interface, which regulates an amount of liquid dispensed from the liquid dispensing system.

Abstract: The present invention relates to an apparatus for the parallel alignment of macromolecules on the surface S of a solid support by displacement of a meniscus formed by the triple interface surface S/solution/air. The apparatus includes a container receiving a solution including the macromolecules to be aligned, a means for immersing the solid support into the solution, with the solid support surface configured to anchor macromolecules, and a means for the relative rectilinear displacement of the surface S and the surface of the solution to thereby cause motion of the meniscus and establish the parallel alignment of the macromolecules. The present invention also relates to a method for the parallel alignment of the macromolecules using the apparatus of the invention.

Abstract: A method for determining the presence, in a liquid medium, of a plurality of different nucleic ligands in the free state comprises: (a) providing a support for plural complexing on which are distributed a plurality of immobilizing sites in which are immobilized a plurality of different nucleic anti-ligands; and (b) contacting the liquid medium with the support, whereby the nucleic ligands are paired with the nucleic anti-ligands. The stability of each ligand/anti-ligand complex is dependent upon at least one exogenous parameter, namely temperature, conditioning the support; the immobilizing sites on the support are differentiated, depending on the exogenous parameter, having respectively different values in the sites; and the anti-ligands are distributed among the different immobilizing sites, depending on the optimum values of the exogenous parameter, determining the stability of the different nucleic ligand/anti-ligand complexes respectively.

Abstract: Methods to determine TIEF-1 polypeptide, or genetic rearrangements in the TIEF-1 gene, in a sample are provided. Also provided are TIEF-1 specific antibodies and nucleic acid probes, and methods of employing those antibodies and probes to detect breast cancer in situ in a mammalian tissue sample.