Applied Biosystems (Life Technologies) has been awarded US Patent No. 7,848,892, "Automatic threshold setting for quantitative polymerase chain reaction."
Lesley Ward, Adrian Jensen, Justin Lyon, Cameron McLeman, and Bryan Tysinger are named as inventors on the patent.
Discloses systems and methods for identifying and quantitating the presence of one or more DNA species in a sample population through PCR amplification. DNA species quantitation includes determining a threshold fluorescence value used to assess the PCR amplification reaction. Various embodiments of the invention incorporate an enhancement function useful in selecting appropriate threshold fluorescence values, facilitating the determination of DNA concentrations by quantitative PCR based methodologies.
Life Technologies has also been awarded US Patent No. 7,847,076, "Methods and reagents for combined PCR amplification."
Paul Mayrand is the sole inventor listed on the patent.
Discloses an oligonucleotide probe that includes an oligonucleotide, a fluorescer molecule attached to a first end of the oligonucleotide, and a quencher molecule attached to the opposite end of the oligonucleotide. The probe is rendered impervious to digestion by the 5'.fwdarw.3' exonuclease activity of a polymerase and the 5'.fwdarw.3' extension by a polymerase. The invention also includes methods for performing combined PCR amplification and hybridization probing. One such method includes the steps of contacting a target nucleic acid sequence with PCR reagents and an oligonucleotide probe as described above, and subjecting these reagents to thermal cycling. One preferred refinement of the method further includes the addition of a strand displacer to facilitate amplification. The patent discloses additional similar combined PCR hybridization methods, such methods not requiring probes having their 5' ends protected, wherein (1) the polymerase lacks 5'.fwdarw.3' exonuclease activity, (2) a 5'.fwdarw.3' exonuclease inhibitor is included, and (3) an exonuclease deactivation step is performed.
Siemens Healthcare Diagnostics has been awarded US Patent No. 7,847,087, "Methods and primers for evaluating HIV-1 mutations."
Tzong-Jyh Huong, Robert Lloyd, and Arejas Uzgiris are named as inventors on the patent.
Describes primer sequences and a method of using such sequences for genotyping HIV-1-containing samples, particularly those which have failed genotyping analysis. The method uses primer sequences designed for analysis of the Group B subtype of the Group M type virus. For example, the method employs a combination of primers, including at least one species of forward primer and at least one species of reverse primer, where the forward primer(s) that can be represented by the degenerate sequence: RARRARGGGCTGYTGGARATGTS; and the reverse primer(s) can be represented by the degenerate sequence: BCHTYACYTTRATCCCSGVRTARATYTGACT or BCHTYACYTTRATCCCSGVRTARATYTGAC. The selected primers, one or more from each group, can be used as reverse transcription, amplification, and sequencing primers, and are suitably packaged in a genotyping kit. Such a kit may include reagents in addition to the primers, such as an RNase inhibitor, a reverse transcriptase, a polymerase, and/or dNTP and ddNTP feedstocks.
NuGen Technologies has been awarded US Patent No. 7,846,733, "Methods and compositions for transcription-based nucleic acid amplification."
Nurith Kurn is the sole inventor listed on the patent.
Describes methods for isothermal exponential amplification of a target polynucleotide. The methods employ two transcription modules. The first module provides linear amplification resulting in RNA transcripts, and the second module provides for further (generally cyclical) amplification resulting in more RNA transcripts. In one aspect, the amplification of the first module is composite-primer based. In a second aspect, the amplification of the first module is based on target switching to generate a primer extension product comprising a promoter sequence. In all aspects, the RNA transcripts of the first transcription module are subjected to further amplification by creating an intermediate product comprising a double-stranded promoter region from which transcription can occur. The invention further provides compositions and kits for practicing said methods, as well as methods which use the amplification results.
NuGen has also been awarded US Patent No. 7,846,666, "Methods of RNA amplification in the presence of DNA."
Nurith Kurn is the sole inventor named on the patent.
Provides methods for amplification of RNA. The methods are particularly suitable for specifically amplifying RNA in the presence of DNA. The methods involve producing a marked first primer extension product from a target RNA in the presence of a DNA-dependent DNA polymerase inhibitor, which prevents replication of DNA by the reverse transcriptase enzyme. The marked nucleic acid products are subsequently selectively amplified in the presence of non-marked nucleic acids. The methods are useful for producing and analyzing polynucleotide sequences complementary to an RNA sequence. The methods are useful for preparing nucleic acid libraries and substrates for analyzing gene expression of cells in biological samples. The invention also provides compositions and kits for practicing the amplification methods, as well as methods that use the amplification products.
Olympus has been awarded US Patent No. 7,846,696, "Method for estimating target nucleic acid ratio."
Tetsuya Tanabe is the sole inventor listed on the patent.
Discloses a method for estimating a ratio between the amount of target nucleic acid and the amount of reference nucleic acid in examined nucleic acid samples from PCR reaction solutions at the endpoint.
Riken has been awarded US Patent No. 7,846,694, "Process for producing template DNA and process for producing protein in cell-free protein synthesis system with the use of the same."
Yoko Motoda, Takashi Yabuki, Takanori Kigawa, and Shigeyuki Yokoyama are named as inventors on the patent.
Describes a method of producing a template DNA used for protein synthesis. The method comprises a step of amplifying a linear double-stranded DNA by PCR using a reaction solution comprising: (1) a first double-stranded DNA fragment comprising a sequence coding for a protein or a portion thereof; (2) a second double-stranded DNA fragment comprising a sequence overlapping with the 5' terminal region of the first DNA fragment; (3) a third double-stranded DNA fragment comprising a sequence overlapping with the 3' terminal region of the first DNA fragment; (4) a sense primer that anneals with the 5' terminal region of the second DNA fragment; (5) and an anti-sense primer that anneals with the 3' terminal region of the third DNA fragment. The second DNA fragment comprises a regulatory sequence for transcription and translation of a gene, and the concentrations of the second DNA fragment and the third DNA fragment in the reaction solution each range from 5 to 2,500 pmol/L. The method enables efficient production of a template DNA for expression and purification of a protein, according to the patent's abstract.
Hologic has been awarded US Patent No. 7,846,661, "Methods of detecting an amplified nucleic acid."
Joseph Sorge is the sole inventor listed on the patent.
The invention relates to methods of generating a signal indicative of the presence of a target nucleic acid sequence in a sample. The methods comprise incubating a sample comprising an amplified target nucleic acid and a nucleic acid polymerase that substantially lacks 5' to 3' exonuclease activity; adding a thermostable FEN nuclease consisting of 5' to 3' exonuclease and/or endonuclease activity so as to cleave a cleavage structure and generate a signal.
Qiagen has been awarded US Patent No. 7,846,315, "Integrated bio-analysis and sample preparation system."
Varouj Amirkhanian and Ming-Sun Lin are named as inventors on the patent.
Describes an integrated bio-analysis system that incorporates built-in sample preparation capabilities. In one aspect, the instrument has a built-in sample preparation device based on PCR (or thermal cycling block/module). In one embodiment, a Peltier unit in the sample preparation device provides thermal cycling of samples supported in a multi-well tray. In another aspect, a [capillary electrophoresis] instrument has a built-in sample preparation capability, which may comprise a sample preparation (bio-molecular reaction) device based on thermal cycler type. In another aspect, a PCR device may have a built-in analysis device, such as a [capillary electrophoresis] device, for verifying the results of the PCR process.