Sequencing-Related US Patents Granted Dec. 23, 2009 — Jan. 26, 2010
US Patent 7,648,824. Method of sequencing DNA
Inventors: Pal Nyren, Mostafa Ronaghi, Annika Tallsjo
Provides a method for identifying a base at a target position in a sample nucleic acid sequence. A primer is hybridized to a sample nucleic acid immediately adjacent to the target position. Next, a polymerase primer extension reaction takes place in the presence of a nucleotide, which will only be incorporated if it is complementary to the base in the target position. The method then determines whether the nucleotide is incorporated by detecting whether PPi is released.
US Patent 7,645,866. Methods of producing and sequencing modified polynucleotides
Inventors: Douglas Smith, Kevin McKernan
Assignee: Life Technologies
Encompasses methods for producing a modified polynucleotide sequence that comprises a phosphorothiolate linkage, methods for determining such a polynucleotide sequence, and methods for separating forward and reverse extension products. The invention also includes kits for producing and/or determining the sequence of a modified polynucleotide that comprises a phosphorothiolate linkage.
US Patent 7,645,596. Method of determining the nucleotide sequence of oligonucleotides and DNA molecules
Inventors: Peter Williams, Thomas Taylor, Daniel Williams, Ian Gould, Mark Hayes
Assignee: Arizona Board of Regents
The invention relates to a novel method for analyzing nucleic acid sequences based on real-time detection of DNA polymerase-catalyzed incorporation of each of the four nucleotide bases, supplied individually and serially in a microfluidic system to a reaction cell containing a DNA template and an oligonucleotide primer. Incorporation of a nucleotide base into the template can be detected by any of a variety of methods, including fluorescence and chemiluminescence detection. Alternatively, microcalorimetic detection of the heat generated by the incorporation of a nucleotide using thermopile, thermistor, and refractive index measurements can be used to detect extension reactions.
US Patent 7,642,344. Methods and compositions for determining the sequence of nucleic acid molecules
Inventors: Jeffrey Van Ness, John Tabone, Jeffry Howbert, John Mulligan
Assignee: Operon Biotechnologies
Provides methods and compounds for determining the sequence of nucleic acid molecules, including multiple nucleic acid sequences simultaneously. The compounds are used to generate tagged nucleic acid fragments that are complementary to a selected target nucleic acid molecule. Each tag is correlative with a particular nucleotide and, in a preferred embodiment, can be detected by mass spectrometry. Following separation of the tagged fragments by sequential length, the tags are cleaved off and are detected by mass spectrometry in order to determine the sequence of the nucleic acid molecule. The individual steps of the methods can be used in automated format.
US Patent 7,638,034. Electrochemical detection of single molecules using abiotic nanopores having electrically tunable dimensions
Inventors: Jose-Maria Sansinena, Antonio Redondo, Virginia Olazabal, Mark Hoffbauer, Elshan Akhadov
Assignee: Los Alamos National Security
Describes a barrier structure for use in an electrochemical stochastic membrane sensor for single molecule detection. The sensor is based on inorganic nanopores with electrically tunable dimensions. The inorganic nanopores are formed from inorganic materials and an electrically conductive polymer. Methods of making the barrier structure and using it to sense single molecules are also described.
US Patent 7,638,276. In vitro sorting method
Inventors: Andrew Griffiths, Dan Tawfik
Assignee: 454 Life Sciences
Describes a method for isolating one or more genetic elements encoding a gene product with a desired activity. The method involves compartmentalizing genetic elements into microcapsules; expressing the genetic elements to produce their respective gene products within the microcapsules; and sorting the genetic elements that produce the gene product with the desired activity. The invention enables the in vitro evolution of nucleic acids by repeated mutagenesis and iterative applications of the method.