Abstract: The present invention relates generally to a method to reduce, substantially reduce, inactivate or eliminate adventitious agents and/or toxins in a sample, particuarly in nutritive media, media supplements, media subgroups and buffer formulations. Specifically, the present invention provides powdered nutritive media, media supplements and media subgroups produced by the methods of the invention, particuarly cell culture media supplements (including powdered sera such as powdered fetal bovine serum (FBS)). The invention further provides powdered buffer formulations produced by the methods of the invention. The invention also provides kits and methods for cultivation of prokaryotic and eukaryotic cells, particularly bacterial cells, yeast cells, plant cells and animal cells (including human cells) using these nutritive media, media supplements, media subgroups and buffer formulations.

Abstract: Recombinational cloning is provided by the use of nucleic acids, vectors and methods, in vitro and in vivo, for moving or exchanging segments of DNA molecules using engineered recombination sites and recombination proteins to provide chimeric DNA molecules that have the desired characteristic(s) and/or DNA segment(s).

Abstract: An improved method is disclosed for diagnosing the presence of a chromosomal translocation characteristic of acute myelogenous leukemia. Nucleic acid molecules that may be used in this improved method are described.

Abstract: A system, method and computer program product is provided for processing images of an electrophoretic separation medium to determine the unit activity of an enzyme. A test aliquot, comprising a macromolecule (such as, DNA, RNA, protein, peptide or the like) and diluted enzyme concentration, is distributed in the separation medium. The enzyme concentration acts as a catalysis to cleave the macromolecule into distinct fragments during electrophoresis. A set of intensity data profiles are produced from images of the fragments. The profiles are stacked and vertically aligned to designate and assign the fragments to their respective lanes. A group of partial bands and final bands are selected from the fragments. Peak integrations are implemented to measure the intensity of the partial and final bands. A series of intensity ratios are computed from the peak integrations. The intensity ratios are normalized and plotted to produce a trend.

Abstract: The invention is related generally to methods of amplifying or synthesizing or producing nucleic acid molecules using Translesion DNA polymerases. In particular, the invention relates to methods of introducing a random mutation into a nucleic acid and encoded polypeptide using Translesion DNA polymerases. The invention also relates to methods of introducing a modified nucleotide into a nucleic acid using Translesion DNA polymerases. The invention also relates to mutagenized and modified nucleic acid molecules and proteins produced by these methods, and to fragments or derivatives thereof. The invention also relates to vectors and host cells comprising mutagenized nucleic acid molecules, fragments, or derivatives. The invention also relates to the use of mutagenized nucleic acid molecules to produce desired polypeptides and uses of modified nucleic acid molecules to analyze samples. The invention also relates to kits or compositions or compounds for use in the invention or for carrying out the invention.

Abstract: The present invention provides novel primers and methods for the detection of specific nucleic acid sequences. The primers and methods of the invention are useful in a wide variety of molecular biology applications and are particularly useful in allele specific PCR.

Abstract: The invention relates to a substantially pure thermostable DNA polymerase from Thermotoga (Tne and Tma) and mutants thereof. The Tne DNA polymerase has a molecular weight of about 100 kilodaltons and is more thermostable than Taq DNA polymerase. The mutant DNA polymerase has at least one mutation selected from the group consisting of (1) a first mutation that substantially reduces or eliminates 3′→5′ exonuclease activity of said DNA polymerase; (2) a second mutation that substantially reduces or eliminates 5′→3′ exonuclease activity of said DNA polymerase; (3) a third mutation in the O helix of said DNA polymerase resulting in said DNA polymerase becoming non-discriminating against dideoxynucleotides. The present invention also relates to the cloning and expression of the wild type or mutant DNA polymerases in E. coli, to DNA molecules containing the cloned gene, and to host cells which express said genes.

Abstract: The present invention provides improved methods, compositions and kits for amplifying a nucleic acid molecule. Specifically, the invention involves replacing at least one nucleotide of an oligonucleotide with nucleotide residue having altered base-pairing characteristics, such as inosine, hypoxanthine, xanthine, and methylated nucleotide derivatives, so as to more equalize the efficiency with which that oligonucleotide and a second oligonucleotide hybridize to a target molecule, and then amplifying the target molecule using, for example, the polymerase chain reaction. Improved amplification results from the improvement in the relative hybridization efficiencies.

Abstract: The invention relates to a gene which encodes reverse transcriptase having DNA polymerase activity and substantially no RNase H activity. The invention also relates to vectors containing the gene and hosts transformed with the vectors of the invention. The invention also relates to a method of producing reverse transcriptase having DNA polymerase activity and substantially no RNase H activity by expressing the reverse transcriptase genes of the present invention in a host. The invention also relates to a method of producing cDNA from mRNA using the reverse transcriptase of the invention. The invention also relates to a kit for the preparation of cDNA from mRNA comprising the reverse transcriptase of the invention.

Abstract: Disclosed are methods and compositions for detecting a chromosomal rearrangement in a sample of nucleic acids. In an exemplary method, a first nucleic acid comprising a portion of a first chromosome, which may be detectably labeled, is attached to a substrate; a first portion of a test nucleic acid is hybridized to the first nucleic acid; a second nucleic acid comprising all or a portion of a second chromosome, which may be detectably labeled, is hybridized to a second portion of the test nucleic acid, thereby forming a trimolecular sandwich, and the hybridization of the test nucleic acid to both of the first and second nucleic acids is detected as an indication that the test nucleic acid comprises a chromosomal rearrangement. In particular embodiments, the first and second nucleic acids are derived from the same chromosome.

Abstract: The present invention relates generally to compositions and methods for enhancing recombinational cloning of nucleic acid molecules. In particular, the invention relates to compositions comprising one or more ribosomal proteins and one or more additional protein components required for recombinational cloning. More particularly, the invention relates to such compositions wherein the ribosomal proteins are one or more E. coli ribosomal proteins, still more particularly wherein the ribosomal proteins are selected from the group of E. coli ribosomal proteins consisting of S10, S14, S15, S16, S17, S18, S19, S20, S21, L20, L21, and L23 through L34, and most particularly S20, L27, and S15. The invention also relates to the use of these compositions in methods for recombinational cloning of nucleic acids, in vitro and in vivo, to provide chimeric DNA molecules that have particular characteristics and/or DNA segments.

Abstract: The present invention provides improved methods for amplifying a nucleic acid molecule. More specifically, the invention provides methods for nucleic acid amplification which use primers having equivalent priming efficiency.

Abstract: The present invention relates generally to nutritive medium, medium supplement, media subgroup and buffer formulations that contain lipid. Specifically, the present invention provides powder nutritive medium, medium supplement and medium subgroup formulations, particularly cell culture medium supplements (including powdered sera such as powdered fetal bovine serum (FBS)), medium subgroup formulations and cell culture media comprising all of the necessary nutritive factors including lipid components that facilitate the in vitro cultivation of cells. The invention is particularly directed to methods of production of these media, media supplement, media subgroup and buffer formulations, and also provides kits and methods for cultivation of prokaryotic and eukaryotic cells, particularly bacterial cells, yeast cells, plant cells and animal cells (including human cells) using these dry powder nutritive media, media supplement, media subgroup and buffer formulations.

Abstract: The present invention is a cell-free subcloning system utilizing three elements: (1) a donor vector that contains a nucleic acid sequence to be transferred to another vector flanked by a site-specific recombination sequence and one or more optional additional nucleic acid sequences, (2) an acceptor vector that contains a site-specific recombination sequence and one or more optional additional nucleic acid sequences, and (3) a site-specific recombinase that recognizes the site-specific recombination sequences in the donor and acceptor vectors so as to transfer the transfer sequence from the donor to the acceptor vector upon contact of the three elements of the system. Also disclosed are rapid subcloning methods employing the vectors and enzymes disclosed herein and kits for use in such methods.

Abstract: The invention described herein comprises libraries of expressible gene sequences. Such gene sequences are contained on plasmid vectors designed to endow the expressed proteins with a number of useful features such as affinity purification tags, epitope tags, and the like. The expression vectors containing such gene sequences can be used to transfect cells for the production of recombinant proteins. A further aspect of the invention comprises methods of identifying binding partners for the products of such expressible gene sequences.

Abstract: The invention relates to a process for deblocking substantially a blocked, detectably labeled oligonucleotide by contacting the blocked detectably labeled oligonucleotide with an effective amount of a nucleophilic amino compound under conditions that result in substantial deblocking of the oligonucleotide, thereby giving the substantially deblocked oligonucleotide.

Abstract: The present invention relates to polypeptides, compositions and methods for extending nucleic acid molecules. Specifically, the present invention discloses polypeptides able to extend double-stranded and/or single-stranded nucleic acid molecules and/or single-stranded/double-stranded nucleic acid complexes (e.g., primer/template complexes, double-stranded templates, single-stranded templates or single-stranded primers) and more particularly when a mismatched dimer is present at or near the 3′ terminus of the primer. Accordingly, the polypeptide of the invention extends nucleic acids in a non-templated fashion. The invention also relates to kits for synthesizing and extending nucleic acid molecules comprising one or more of the polypeptides or compositions of the invention. The invention also relates to compositions prepared for carrying out the methods of the invention and to compositions made after or during such methods.

Abstract: A gel and buffer system for gel electrophoresis wherein separation occurs at neutral pH.

Abstract: The present invention is generally related to compositions, kits and methods for labeling nucleic acid molecules using reverse transcriptases, preferably multi-subunit reverse transcriptases such as ASLV reverse transcriptases. Specifically, the invention relates to methods, kits and compositions for fluorescently labeling nucleic acid molecules during nucleic acid synthesis. The labeled nucleic acid molecules produced in accordance with the invention are particularly suited as labeled probes for nucleic acid detection and diagnostics.

Abstract: The invention relates to a gene which encodes reverse transcriptase having DNA polymerase activity and substantially no RNase H activity. The invention also relates to vectors containing the gene and hosts transformed with the vectors of the invention. The invention also relates to a method of producing reverse transcriptase having DNA polymerase activity and substantially no RNase H activity by expressing the reverse transcriptase genes of the present invention in a host. The invention also relates to a method of producing cDNA from mRNA using the reverse transcriptase of the invention. The invention also relates to a kit for the preparation of cDNA from mRNA comprising the reverse transcriptase of the invention.