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'.fwdarw.5' exonuclease activity of said DNA polymerase; (2) a second mutation that substantially reduces or eliminates 5'.increment.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: This invention relates to a method of incorporating an exo-sample nucleotide into the amplified product strands resulting from a nucleic acid amplification process. Once the product strands have been obtained and analyzed (e.g., by hybridization, Southern blot, etc.), the exo-sample strands can be selectively destroyed by acting on the incorporated exo-sample nucleotide. Two embodiments are presented. In a first embodiment, the exo-sample nucleotide is incorporated by carrying out the amplification reaction in the presence of an excess of exo-sample nucleotide triphosphate. In a second embodiment, the exo-sample nucleotide is incorporated by carrying out the amplification reaction in the presence of an oligonucleotide which has, as part of its sequence, one or more exo-sample nucleotides.

Abstract: The invention relates to a substantially pure thermostable DNA polymerase from Thermotoga neapolitana (Tne) 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 Tne DNA polymerase has at least one mutation selected from the group consisting of (1) a first mutation that substantially reduces or eliminates 3'.fwdarw.5' exonuclease activity of said DNA polymerase; (2) a second mutation that substantially reduces or eliminates 5'.fwdarw.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 Tne DNA polymerase in E. coli, to DNA molecules containing the cloned gene, and to host cells which express said genes.

Abstract: The invention relates to the cloning of a gene encoding rat liver RI, and its expression in a cellular host. In addition, the invention relates to the successful cloning of a gene encoding porcine liver RI, and its expression in a cellular host. The invention also relates to the cloning and expression of human liver RI, and the cloning and expression of chimeric mammalian RIs, particularly chimeric porcine/human liver RIs, which may be thermostable. The invention also relates to methods and kits for use in producing nucleic acid molecules and polypeptides using the RIs of the invention, and to nucleic acid molecules and polypeptides produced using these methods and kits.

Abstract: The invention relates to a substantially pure thermostable DNA polymerase from Thermotoga neapolitana (Tne). The Tne DNA polymerase has a molecular weight of about 100 kilodaltons and is more thermostable than Taq DNA polymerase. The present invention also relates to the cloning and expression of the Tne DNA polymerase in E. coli, to DNA molecules containing the cloned gene, and to host cells which express said genes. The Tne DNA polymerase of the invention may be used in well-known DNA sequencing and amplification reactions.

Abstract: The invention relates to improved E. coli bacteria with enhanced viability at low temperatures, methods for producing improved bacterial strains capable of enhanced viability at low temperatures, and the isolation and use of genetic material capable of enhancing the viability of bacteria at low temperatures. In addition to the enhanced viability at low temperatures, the bacteria may exhibit enhanced transformation efficiencies after storage at low temperatures. As such, the invention may be used for the insertion of exogenous DNA sequences into the bacteria of the invention.

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: A gel electrophoresis apparatus and a cooling system therefor. The apparatus includes a front panel and a back panel. A gel slab is contained in a gel slab platform between the front and back panels. The gel slab platform extends upwardly to the front edge of an upper buffer reservoir and downwardly to a lower buffer reservoir. A conventional gel mold assembly is inserted into the apparatus and supported by the gel slab platform. A cooling unit is disposed below the lower buffer reservoir of the apparatus. A baffle plate is inserted between the front panel and a back panel of the apparatus. When the cooling unit is actuated, the baffle plate directs a cooling medium through the apparatus and between the front panel and the back panel to cool the gel slab. The apparatus also includes dampers which are pivotally mounted on the apparatus at the top of the baffle plate. The dampers are used to control the flow of the cooling medium through each baffle individually.

Abstract: The present invention provides improved methods for amplifying a nucleic acid molecule. More specifically, the invention involves replacing at least one nucleotide of an oligonucleotide with a nucleotide having altered base pairing characteristics, 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 relative hybridization efficiencies.

Abstract: An automated dispensing device is provided that dispenses a calibrated quantity of a fluid into a receptacle having a plurality of spaced-apart rows of receiving wells. The calibrated quantity of fluid is determined based upon a dispensing time and a dispensing scale factor that accounts for the viscosity of the fluid to be dispensed. The automated dispensing device can be configured with independently controllable nozzles for selective delivery of fluid. The automated dispensing device is particularly suitable for dispensing a calibrated quantity of ammonium hydroxide into the receiving wells of each row of a microtiter plate for oligonucleotide cleavage.

Abstract: The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).

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: The present invention discloses highly packed polycationic ammonium, sulfonium and phosphonium lipid compounds useful for making lipid aggregates for delivery of macromolecules and other compounds into cells. They are especially useful for the DNA-dependent transformation of cells. Methods for their preparation and use as intracellular delivery agents are also disclosed.

Abstract: The invention relates to a nucleic acid marker ladder which is a restriction endonuclease digest, wherein a nucleic acid restriction endonuclease digest is a collection of nucleic acid fragments resulting from complete digestion of one or more nucleic acids by one or more restriction endonucleases; the restriction endonuclease digest contains at least 3 fragments; and the size of the fragments in base pairs is a multiple of an integer, wherein the integer is 10 or more.

Abstract: The present invention discloses the cloning and expression in Escherichia coli of the SstI/SacI restriction-modification system. The SstI methylase gene was cloned by methylase selection (the "Hungarian Trick" and the orientation and boundaries of the gene were established. The level of methylation of the host was improved significantly by expressing the SstI methylase from the strong RPSL promoter. Using the SstI methylase clone as a probe, a chromosomal map was generated in the area of the gene for the SstI methylase. DNA downstream from the methylase was cloned and found to contain the gene for the SstI endonuclease. The orientation of the SstI endonuclease was established and a strong promoter was placed close to the SstI endonuclease gene by generating nested deletions. By these methods, an E. coli strain was constructed which produced high levels of SstI endonuclease. The SacI locus in the S. achromogenes chromosome was found to be identical to the SstI locus in S. stanford in all measured respects.

Abstract: The present invention provides a method for the rapid isolation and recovery of a desired target DNA or RNA molecules from a mixture or library containing such molecules. The method involves the use of biotinylated probes and enzymatic repair-cleavage to eliminate undesired library members from a sample.

Abstract: The present invention discloses compositions useful for transfecting eukaryotic cells comprising nucleic acid complexes with peptides, proteins or protein fragments, wherein the peptide is optionally covalently coupled to a DNA-binding group, and cationic lipids useful for transfecting eukaryotic cells. Methods for the preparation of transfecting compositions and use as intracellular delivery agents and extracellular targeting agents are also disclosed.

Abstract: The present invention discloses a recombinant DNA molecule having a structural gene encoding a processive, thioredoxin-independent DNA polymerase that is substantially reduced in processive 3'-to-5' DNA exonuclease activity, a promoter, and an origin of replication. A method for producing this enzyme is also disclosed, as is the protein produced by this process. This invention is exemplified by expression of exonuclease.sup.- T5 DNA polymerase in E. coli.

Abstract: A cooler assembly for maintaining vials filled with enzyme samples in a chilled state. The cooler assembly includes a foam block having wells disposed therein for holding vials of samples in an upright position. The cooler assembly also includes an outer foam box and a lid. The foam block is positioned inside the outer foam box. The lid is then positioned on top of the outer foam box to provide further insulation for the foam block. In use, the foam block is filled with a liquid, such as water, via a fill hole. The foam block is then frozen. Vials are placed in the wells of the frozen foam block so that the contents of the vials remain cool when placed in a room having an ambient temperature. The inner foam block is made from self-skinning, open cell foam. The skin on the foam block renders the block leakproof. The cooler assembly provides for efficient heat transfer between the chilled wells of the foam block and the contents of the vials.

Abstract: This invention relates to a method of incorporating an exo-sample nucleotide into the amplified product strands resulting from a nucleic acid amplification process. Once the product strands have been obtained and analyzed (e.g., by hybridization, Southern blot, etc.), the exo-sample strands can be selectively destroyed by acting on the incorporated exo-sample nucleotide.Two embodiments are presented. In a first embodiment, the exo-sample nucleotide is incorporated by carrying out the amplification reaction in the presence of an excess of exo-sample nucleotide triphosphate.In a second embodiment, the exo-sample nucleotide is incorporated by carrying out the amplification reaction in the presence of an oligonucleotide which has, as part of its sequence, one or more exo-sample nucleotides.