Abstract: The subject invention provides for a strain of host cells that contains a temperature sensitive variant of the gene encoding the endonuclease VII from phage T4. Using this host strain, the invention features a novel cloning method that selects for PCR products that are devoid of PCR-generated mutations.
Type:
Application
Filed:
June 26, 2002
Publication date:
January 1, 2004
Applicant:
Stratagene
Inventors:
Alan L. Greener, Lisa Joy Hexdall, Carsten-Peter Carstens, Joseph A. Sorge
Abstract: The present invention relates to compositions and kits comprising a mutant DNA polymerase with increased reverse transcriptase activity. The invention also relates to methods for using the subject compositions and kits.
Type:
Application
Filed:
May 12, 2003
Publication date:
December 11, 2003
Applicant:
Stratagene
Inventors:
Bahram Arezi, Holly Hogrefe, Joseph A. Sorge, Connie Jo Hansen
Abstract: An apparatus for thermally cycling samples of a biological material including a thermal block assembly including a plurality of sample holders for receiving samples of biological material; a heat sink thermally coupled to the thermal block assembly, the heat sink transferring heat away from the thermal block assembly to ambient air in contact with the heat sink; a first heat source thermally coupled to the thermal block assembly to provide heat to the thermal block assembly; and a second heat source thermally coupled to the first heat source and configured to provide heat to a portion of the first heat source. The arrangement of the heat sink, first heat source and second heat source can provide substantial temperature uniformity among the plurality of sample holders. The invention also includes a method for thermally cycling samples of biological material.
Abstract: The present invention provides a polynucleotide encoding a green fluorescent protein from Renilla mulleri comprising a humanized sequence which permits enhanced expression of the encoded polypeptide in mammalian cells.
Type:
Grant
Filed:
April 19, 2001
Date of Patent:
November 11, 2003
Assignee:
Stratagene
Inventors:
Joseph A. Sorge, Peter Edward Vaillancourt
Abstract: cDNA microarrays are provided for increasing the accuracy and reliability of expression profiling techniques and for identifying new genes. An array comprises a plurality of nucleic acid members, each member having a unique position and stably associated with a solid support. Each nucleic acid member comprises a noncoding sequence present at either the 3′-end or the 5′-end of an RNA transcript (e.g., such as an untranslated region or UTR). In one embodiment, each nucleic acid member is less than 1000 nucleotides. In another embodiment, each nucleic acid member is less than 600 nucleotides. In a further embodiment of the invention, each nucleic acid member comprises substantially noncoding sequences.
Abstract: The invention provides a method and related kits and reagents for producing, purifying, isolating, or enriching desired nucleic acids from a library. The desired nucleic acids are produced from polymerase enzyme extension products, generated from specific primers or sets of primers, in a form that is immediately replicable in a host cell. The invention can be practiced in solution phase, thus eliminating the need for solid phase filter hybridizations, column hybridizations, or gel electrophoresis purification when enriching for or isolating a target sequence or vector from one or more libraries.
Abstract: This invention provides methods for producing and selecting host cells that better survive transformation treatment by subjecting host cells to conditions that alter them, subjecting the altered cells to transformation conditions, and selecting host cells that survive the transformation conditions. This invention also provides methods for transferring nucleic acids of interest into host cells, using cells that are better able to survive transformation treatment. Also, this invention provides kits for producing or selecting host cells in transformation treatments, as well as, kits comprising various host cells that may be utilized in transformation experiments.
Abstract: A method of producing libraries of genes encoding antigen-combining molecules or antibodies is described. In addition, a method of producing antigen-combining molecules which does not require an in vivo procedure is described. Vectors useful in the present method and antigen-combining molecules produced by the method are discussed. The antigen-combining molecules are useful for the detection, quantitation, purification and neutralization of antigens, as well as for diagnostic, therapeutic and prophylactic purposes.
Abstract: The invention concerns an expression vector that permits expression of genes and fragments thereof in both prokaryotic and mammalian systems. The invention also pertains to derivatives of such vector that contain one or more prokaryotic or eukaryotic (especially mammalian) genes or gene fragments. The invention further pertains to prokaryotic or mammalian cells containing such an expression vector or derivative.
Abstract: The invention relates to compositions comprising a first fusion protein comprising a first polypeptide domain and a R. reniformis luciferase and a second fusion protein comprising a second polypeptide domain and a R. reniformis GFP. The invention also relates to compositions comprising one or more polynucleotides encoding a first fusion protein comprising a first polypeptide domain and a R. reniformis luciferase and a second fusion protein comprising a second polypeptide domain and a R. reniformis GFP. The invention also relates to methods and kits for detecting protein-protein interactions, determining the location of a protein-protein interaction, identifying cells wherein there is a protein-protein interaction of interest, and screening for a candidate modulator that increases or decreases the amount of a protein-protein interaction.
Abstract: This invention is directed to methods and kits for creating and analyzing molecules using uniquely identifiable tags. The invention is also directed to methods and kits that use uniquely identifiable tags for sequencing DNA, for determining mutations, including substitutions, deletions, and additions, in sample genes, and monitoring mRNA populations.
Abstract: The invention relates to compositions and methods for nucleic acid PCR mutagenesis using novel error-prone DNA polymerases and a PCR enhancing factor. The invention also relates to compositions and methods for nucleic mutagenesis with two or more DNA polymerases lacking or exhibiting reduced exonuclease activity. The invention further relates to kit format of said compositions for PCR mutagenesis.
Abstract: The present invention relates to a method for identifying a nucleotide at a predetermined location on a target polynucleotide. The method involves single nucleotide extension reaction comprising an oligonucleotide primer comprising a first sequence and a second sequence or a tag. The method may further comprises a probe which hybridizes to the second sequence or an anti-tag molecule which interacts with the tag, where the hybridization or interaction causes a detectable signal transfer which is indicative of the identity of the nucleotide base at the predetermined location. The invention further provides compositions and kits for performing the subject method of the invention.
Type:
Application
Filed:
January 24, 2002
Publication date:
August 7, 2003
Applicant:
Stratagene
Inventors:
Joseph A. Sorge, Bahram Arezi, Holly Hogrefe
Abstract: The invention relates to a method of generating a signal indicative of the presence of a target nucleic acid sequence in a sample, where the method includes forming a cleavage structure by incubating a sample comprising a target nucleic acid sequence with a probe having a secondary structure that changes upon binding of the probe to the target nucleic acid sequence, and cleaving the cleavage structure with a nuclease to release a nucleic acid fragment to generate a signal, wherein generation of the signal is indicative of the presence of a target nucleic acid sequence in a sample.
Abstract: This invention provides improved electroporation methods for transferring nucleic acids of interest into host cells, wherein the host cells are (1) suspended in a substantially non-ionic solution comprising at least one sugar or sugar derivative, (2) mixed with the nucleic acids of interest, and (3) electrically treated. Also, this invention provides for kits used in the method for transferring nucleic acids into host cells.
Abstract: The detection of insertions and/or deletions in reiterated nucleotide sequences in tissues provides an identification of neoplastic changes that are associated with malignancy. The mutations are preferably detected by PCR based amplification of target sequences using selected primers, followed by standard analytic procedures. The detection of these mutations is useful as a diagnostic tool for cancer development and has direct application for cancer prognosis.
Type:
Grant
Filed:
December 27, 1995
Date of Patent:
May 20, 2003
Assignee:
Stratagene
Inventors:
Manuel Perucho, Miguel Angel Peinado, Yurij Ionov, Sergei Malkhosyan, Michael McClelland, John Welsh
Abstract: The invention relates to fluorescent dyes. More particularly, the invention relates to fluorescent cyanine dyes, and especially to water soluble fluorescent cyanine dyes that contain additional sites for attachment to biomolecules. The invention provides a group of novel, water soluble fluorescent cyanine dyes that have distinct fluorescence characteristics that permit their use in any assay or method suited to water soluble fluorescent dyes, and especially to assays requiring a plurality of distinguishable fluorescent markers. The invention further relates to nucleotides, nucleosides, polynucleotides and polypeptides labeled with novel fluorescent cyanine dyes according to the invention, and methods of using them.
Abstract: The invention relates to a method of generating a signal indicative of the presence of a target nucleic acid sequence in a sample, where the method includes forming a cleavage structure by incubating a sample containing a target nucleic acid sequence with a nucleic acid polymerase and cleaving the cleavage structure with a FEN nuclease to generate a cleaved nucleic acid fragment. The invention also relates to methods of detecting or measuring a target nucleic acid sequence, where the method includes forming a cleavage structure by incubating a target nucleic acid sequence with a nucleic acid polymerase, cleaving the cleavage structure with a FEN nuclease and detecting or measuring the release of a fragment.
Abstract: The invention provides improved methods of introducing site-directed mutations into circular DNA molecules of interest by means of mutagenic primer pairs. The mutagenic primer pairs are also selected so as to be either completely complementary or partially complementary to each other, wherein the mutation site (or sites) is located within the region of complementarity. A mutagenic primer pair is annealed to opposite strands of a circular DNA molecule containing the DNA sequence to be mutagenized. After annealing, first, and second mutagenized DNA strands, each incorporating a member of the mutagenic oligonucleotide primer pair is synthesized by a linear cyclic amplification reaction. After the linear cyclic amplification mediated synthesis step is completed, the reaction mixture is treated with a selection enzyme that digests the parental template strands. After the digesting step, a double-stranded circular DNA intermediate is formed.
Type:
Application
Filed:
August 26, 2002
Publication date:
April 3, 2003
Applicant:
Stratagene and Children's Medical Center Corporation
Inventors:
John C. Bauer, Dowain A. Wright, Jeffrey Carl Braman, Raif S. Geha
Abstract: The invention relates to a method of generating a signal indicative of the presence of a target nucleic acid sequence in a sample, where the method includes forming a cleavage structure by incubating a sample containing a target nucleic acid sequence with a nucleic acid polymerase and cleaving the cleavage structure with a FEN nuclease to generate a cleaved nucleic acid fragment. The invention also relates to methods of detecting or measuring a target nucleic acid sequence, where the method includes forming a cleavage structure by incubating a target nucleic acid sequence with a nucleic acid polymerase, cleaving the cleavage structure with a FEN nuclease and detecting or measuring the release of a fragment.