Abstract: Positive reference spiked in collected sample for use in qualitatively and quantitatively detecting viral RNA.

Abstract: The present invention discloses a reference DNA and the use thereof, wherein the reference DNA is selected from the group consisting of: (i) DNA fragment 1: characterized in that it carries a defined gene mutation and at least one another artificially altered base X2, wherein, as compared to a wild type of the gene, at least one defined base X1 in the defined gene mutation undergoes a mutation associated with the occurrence, diagnosis, and/or treatment (such as a target targeted by a medicament) of a disease (such as a tumor), wherein the mutation is a substitution mutation, a deletion mutation, and/or an insertion mutation, and the artificially altered base X2 is different from the mutant base X1 which is contained in the DNA of a sample to be detected and defined to be associated with the occurrence, diagnosis and/or treatment of a disease, (ii) DNA fragment 2: characterized in that it differs from the DNA fragment 1 only in that it does not comprise the defined base X1 mutation, or (iii) a mixture of th

Abstract: The present invention relates to a DNA and RNA polymerases which have increased fidelity (or reduced misincorporation rate). In particular, the invention relates to a method of making such polymerases by modifying or mutating the nucleotide binding domain of the polymerase (e.g., the O-helix). The invention also relates to DNA molecules containing the genes encoding the polymerases of the invention, to host cells containing such DNA molecules and to methods to make the polymerases using the host cells. The polymerases are particularly suited for nucleic acid synthesis, sequencing, amplification and cDNA synthesis.

Abstract: The present invention provides methods for use in identifying, analyzing and typing polymorphic DNA fragments, particularly minisatellite, microsatellite or STR DNA fragments. In particular, the invention provides methods using DNA polymerases, more particularly thermostable DNA polymerases, and most particularly Thermotoga polymerases or mutants or derivatives thereof, whereby minisatellite, microsatellite or STR DNA molecules may be amplified and analyzed for polymorphisms. The invention also relates to polymerases having reduced, substantially reduced or eliminated ability to add non-template 3? nucleotides to a synthesized nucleic acid molecule. In accordance with the invention, such reduction or elimination may be accomplished by modifying or mutating the desired polymerase.

Abstract: Recombinant DNA vectors and methods for cloning and expressing nucleic acid molecules by using a combination of site-specific recombination and end-to-end joining or linking of nucleic acid molecules, such as endonuclease restriction digestion and ligation. The DNAs, vectors and methods can be used for inserting, exchanging, transferring a variety of DNA segment(s) both in vitro and in vivo. Also disclosed are linker molecules and methods using these linkers the can be used for cloning a gene of interest into an expression vector in one-step. The linker sequences comprise adapter sequences for cloning purposes, as well as eukaryotic and prokaryotic ribosome binding sites for increase translation efficiency.

Abstract: A method for removing extraneous nucleotides in a cloned coding sequence using a type IIs endonuclease, the method comprising introducing a linker that comprises at least one recognition site for a Type IIs restriction endonuclease to the ORF, cloning the ORF into a suitable vector, and removing the extraneous nucleotides from the vector with a RE IIs digestion. Also provided are a vector, a kit and oligonucloetides suitable for the invention.

Abstract: Recombinant DNA vectors and methods for cloning and expressing nucleic acid molecules by using a combination of site-specific recombination and end-to-end joining or linking of nucleic acid molecules, such as endonuclease restriction digestion and ligation. The DNAs, vectors and methods can be used for inserting, exchanging, transferring a variety of DNA segment(s) both in vitro and in vivo. Also disclosed are linker molecules and methods using these linkers the can be used for cloning a gene of interest into an expression vector in one-step. The linker sequences comprise adapter sequences for cloning purposes, as well as eukaryotic and prokaryotic ribosome binding sites for increase translation efficiency.

Abstract: The sensitivity and specificity of polynucleotide synthesis is increased by protecting the 3′-end of an oligonucleotide used as a primer in the synthesis of the polynucleotide. Protection of the 3′-end of an oligonucleotide prevents non-specific chain elongation. Removal of blocking group an elevated temperature, using a thermostable enzyme, permits template-specific polynucleotide synthesis. The present invention also provides oligonucleotides with a 3′ end protected by a blocking group and a thermostable enzyme capable of removing the blocking group at an elevated temperature. The compositions and methods of the invention are very useful in a variety of techniques for DNA/RNA amplification and analysis, including medical genetics research and diagnosis, pathogen detection, forensic, and animal and plant genetics applications, among others.

Abstract: The sensitivity and specificity of polynucleotide synthesis is increased by protecting the 3′-end of an oligonucleotide used as a primer in the synthesis of the polynucleotide. Protection of the 3′-end of an oligonucleotide prevents non-specific chain elongation. Removal of blocking group an elevated temperature, using a thermostable enzyme, permits template-specific polynucleotide synthesis. The present invention also provides oligonucleotides with a 3′ end protected by a blocking group and a thermostable enzyme capable of removing the blocking group at an elevated temperature. The compositions and methods of the invention are very useful in a variety of techniques for DNA/RNA amplification and analysis, including medical genetics research and diagnosis, pathogen detection, forensic, and animal and plant genetics applications, among others.

Abstract: The present invention provides methods for use in identifying, analyzing and typing polymorphic DNA fragments, particularly minisatellite, microsatellite or STR DNA fragments. In particular, the invention provides methods using DNA polymerases, more particularly thermostable DNA polymerases, and most particularly Thermotoga polymerases or mutants or derivatives thereof, whereby minisatellite, microsatellite or STRDNA molecules may be amplified and analyzed for polymorphisms. The invention also relates to polymerases having reduced, substantially reduced or eliminated ability to add non-template 3′ nucleotides to a synthesized nucleic acid molecule. In accordance with the invention, such reduction or elimination may be accomplished by modifying or mutating the desired polymerase.

Abstract: The present invention provides methods for use in identifying, analyzing and typing polymorphic DNA fragments, particularly minisatellite, microsatellite or STR DNA fragments. In particular, the invention provides methods using DNA polymerases, more particularly thermostable DNA polymerases, and most particularly Thermotoga polymerases or mutants or derivatives thereof, whereby minisatellite, microsatellite or STR DNA molecules maybe amplified and analyzed for polymorphisms. The invention also relates to polymerases having reduced, substantially reduced or eliminated ability to add non-template 3′ nucleotides to a synthesized nucleic acid molecule. In accordance with the invention, such reduction or elimination may be accomplished by modifying or mutating the desired polymerase.

Abstract: The sensitivity and specificity of polynucleotide synthesis is increased by protecting the 3′-end of an oligonucleotide used as a primer in the synthesis of the polynucleotide. Protection of the 3′-end of an oligonucleotide prevents non-specific chain elongation. Removal of blocking group an elevated temperature, using a thermostable enzyme, permits template-specific polynucleotide synthesis. The present invention also provides oligonucleotides with a 3′ end protected by a blocking group and a thermostable enzyme capable of removing the blocking group at an elevated temperature. The compositions and methods of the invention are very useful in a variety of techniques for DNA/RNA amplification and analysis, including medical genetics research and diagnosis, pathogen detection, forensic, and animal and plant genetics applications, among others.