Abstract: A method is provided for genotyping a target sequence at at least two allelic sites by a 5? nuclease amplification reaction. In one embodiment, the method includes performing a nucleic acid amplification on a target sequence having at least two different allelic sites using a nucleic acid polymerase having 5?-3? nuclease activity and a primer capable of hybridizing to the target sequence in the presence of two or more sets of allelic oligonucleotide probes wherein at least all but one of the allelic oligonucleotide probes include a different fluorescer than the other probes and a quencher positioned on the probe to quench the fluorescence of the fluorescer; detecting a fluorescence spectrum of the amplification; calculating a fluorescence contribution of each fluorescer to the fluorescence spectrum; and determining a presence or absence of the different allelic variants based on the fluorescence contribution of each fluorescer to the combined fluorescence spectrum.
Abstract: Extended rhodamine compounds exhibiting favorable fluorescence characteristics having the structure are disclosed. In addition, novel intermediates for synthesis of these dyes are disclosed, such intermediates having the structure In addition, methods of making and using the dyes as fluorescent labels are disclosed.
Type:
Grant
Filed:
July 29, 2010
Date of Patent:
February 28, 2012
Assignee:
Applied Biosystems, LLC
Inventors:
Joe Y. L. Lam, Scott C. Benson, Steven M. Menchen
Abstract: The invention is directed to a method and device for simultaneously testing a sample for the presence, absence, and/or amounts of one or more of a plurality of selected analytes. The invention includes, in one aspect, a device for detecting or quantitating a plurality of different analytes in a liquid sample. Each chamber may include an analyte-specific reagent effective to react with a selected analyte that may be present in the sample, and detection means for detecting the signal. Also disclosed are methods utilizing the device.
Type:
Grant
Filed:
April 7, 2006
Date of Patent:
February 21, 2012
Assignee:
Applied Biosystems, LLC
Inventors:
Timothy M. Woudenberg, Michael Albin, Reid B. Kowallis, Yefim Raysberg, Robert P. Ragusa, Emily S. Winn-Deen
Abstract: System and method for fluorescent light excitation and detection from samples to enhance the numerical aperture and/or reduce the cross-talk of the fluorescent light.
Abstract: Methods and devices are described for concentration and cleanup of samples containing bio-molecule analytes (e.g., polynucleotides, such as DNA, RNA, PNA). Various embodiments provide for pH-mediated sample concentration and cleanup of nucleic acid samples with channel devices (e.g., cross-T format, microchannel devices).
Abstract: The invention relates to a method for simultaneous quantification of human nuclear DNA and human male DNA in a biological sample while also detecting the presence of PCR inhibitors in a single reaction. The multiplex quantification method also provides a ratio of human nuclear and male DNA present in a biological sample. Such sample characterization is useful for achieving efficient and accurate results in downstream molecular techniques such as genotyping.
Type:
Application
Filed:
August 26, 2011
Publication date:
January 19, 2012
Applicant:
Applied Biosystems, LLC
Inventors:
Jaiprakash G. SHEWALE, Manohar Furtado, Pius Brzoska, Maura Barbisin, Rixun Fang, Michael Malicdem, Cristin O'Shea
Abstract: The synthesis of capped/tagged RNA, methods of use and kits providing same are contemplated. Tagged RNA permits isolation of RNA transcripts in vitro. The ability to isolate and purify capped RNA results in improved transcription and translation and provides a tool for identifying RNA-protein interactions. Such capped RNA finds use in therapeutic applications, diagnosis and prognosis and in the treatment of cancers and HIV.
Type:
Grant
Filed:
April 30, 2010
Date of Patent:
January 10, 2012
Assignee:
Applied Biosystems, LLC
Inventors:
Anilkumar Kore, Shanmugasundaram Muthian, Irudaya Charles
Abstract: The present teachings relate to microfluidic valves and pumping systems, which may be suitable for controlling and facilitating liquid flow. Electrodes are disposed proximately to volumes containing a liquid. The liquid flow can be facilitated by electrowetting forces. Processes for controlling the flow of liquids, as well as for pumping liquids, are also disclosed.
Abstract: Methods for normalizing output from an instrument employing a reference standard or non-fluorescing substance disposed within at least one of a plurality of reaction chambers. The method comprises collecting and analyzing a signal associated with the reference standard or non-fluorescing substance to determine a normalizing bias. The normalizing bias is then applied to the data signal collected from a remainder of the plurality of reaction chambers.
Type:
Grant
Filed:
August 18, 2009
Date of Patent:
January 3, 2012
Assignee:
Applied Biosystems LLC
Inventors:
H. Pin Kao, Ian A. Harding, Achim Karger, Mark F. Oldham, Omead Ostadan, Greg Young
Abstract: The present invention provides methods and systems for an automated method of identifying allele values from data files derived from processed fluorophore emissions detected during the observation of fluorophore labeled nucleotide probes used in analyzing polymorphic DNA are provided. These methods are used in the rapid and efficient distinguishing of targeted polymorphic DNA sites without control samples.
Type:
Grant
Filed:
July 7, 2010
Date of Patent:
January 3, 2012
Assignee:
Applied Biosystems, LLC
Inventors:
Stephen A. Glanowski, Jeremy Heil, Emily S. Winn-Deen, Ivy A. McMullen
Abstract: A method is disclosed for effecting the concentration of a polar analyte in an alternating electric field. In the method, a relative translation of the polar analyte and an alternating electric field along a translation path is effected. A portion of the polar analyte is then trapped and concentrated in a concentration zone formed by the intersection of the translation path and the alternating electric field. Also disclosed are various devices for carrying out the forgoing method.
Abstract: Implementations of the present invention describe an apparatus for generating calibration factors for a spectral detector instrument. The calibration factors are derived from a calibration plate containing one or more spectral species in each well of the calibration plate. Each well is then exposed to an excitation source that causes the one or more spectral species in each of the wells to fluoresce. The signal response is measured and associated with each spectral species at each different well position in the calibration plate. Next, the measured signal response from each spectral species at each well position in the calibration plate is compared with a predetermined signal response for each spectral species. The results of this comparison can be used to determine a calibration factor for each well and spectral species to compensate for the difference between the measured signal response and the predetermined signal response.
Abstract: A microfluidic device comprising a first surface portion and a first sample retaining element, which have differing affinities to a fluid, and a method comprising supplying a sample to such a device. In some embodiments, the differing affinity is a result of plasma, ion embedding, surface charging, chemical, optical, electronic and/or electromagnetic treatment. Also, a microfluidic device comprising at least one microcapillary device having a sample retaining element, at least one surface of which exhibits hydrophobicity, hydrophilicity, electromagnetic force exertion and electrostatic force exertion. Also, a microfluidic device comprising a first element having a hydrophilic pattern comprising at least a first sample retaining element. Also, a method comprising supplying a sample to a channel between a first element and a second element, and inducing in the first element at least one hydrophilic pattern by electrets or by internal or external electrodes to provide a charged surface.
Type:
Grant
Filed:
August 13, 2007
Date of Patent:
November 29, 2011
Assignees:
Applied Biosystems, LLC, The United States of America as represented by the Department of Health and Human Services
Abstract: Binary probe and clamp compositions conduct methods for target hybridization detection. Where the probe is a substrate for exonuclease cleavage, the composition provides quantitation and detection of PCR products, by real-time and end-point measurements. Where the probe is an amplification primer, the composition provides an improved method for labelling and detection of PCR products. Probes and clamps may be labelled with fluorescent dyes, quenchers, hybridization-stabilizing moieties, chemiluminescent dyes, and affinity ligands. Clamps may be nucleic acid analogs, such as 2-aminoethylglycine PNA.
Type:
Grant
Filed:
August 10, 2009
Date of Patent:
November 29, 2011
Assignee:
Applied Biosystems, LLC
Inventors:
Kenneth Livak, Michael Egholm, Michael Hunkapiller
Abstract: The invention is directed to a method and device for simultaneously testing a sample for the presence, absence, and/or amounts of one or more of a plurality of selected analytes. The invention includes, in one aspect, a device for detecting or quantitating a plurality of different analytes in a liquid sample. Each chamber may include an analyte-specific reagent effective to react with a selected analyte that may be present in the sample, and detection means for detecting the signal. Also disclosed are methods utilizing the device.
Type:
Grant
Filed:
May 10, 2006
Date of Patent:
November 29, 2011
Assignee:
Applied Biosystems, LLC
Inventors:
Timothy M. Woudenberg, Michael Albin, Reid B. Kowallis, Yefim Raysberg, Robert P. Ragusa, Emily S. Winn-Deen
Abstract: Methods of detecting at least one analyte and at least one nucleic acid in a sample are provided. Reagents for carrying out the methods are also provided.
Abstract: A fluid processing device is provided that includes a substrate, a plurality of fluid retainment regions formed in or on the substrate, and a barrier at least partially separating two or more of the fluid retainment regions. The barrier includes a mixture of a sequestering material and a reaction component. The reaction component can be at least one of a reactant, a reagent, a catalyst, an initiator, a promoter, a cofactor, an enzyme, a salt, or a combination thereof. The sequestering material can be a porous material, a dissolvable material, or both.
Type:
Grant
Filed:
October 18, 2005
Date of Patent:
November 22, 2011
Assignee:
Applied Biosystems, LLC
Inventors:
Konrad Faulstich, Aldrich N. K. Lau, Debjyoti Banerjee, Umberto Ulmanella, Jun Xie
Abstract: The invention is directed to a method and device for simultaneously testing a sample for the presence, absence, and/or amounts of one or more a plurality of selected analytes. The invention includes, in one aspect, a device for detecting or quantitating a plurality of different analytes in a liquid sample. The device includes a substrate which defines a sample-distribution network having (i) a sample inlet, (ii) one or more detection chambers, and (iii) channel means providing a dead-end fluid connection between each of the chambers and the inlet Each chamber may include an analyte-specific reagent effective to react with a selected analyte that may be present in the sample, and detection means for detecting the signal. Also disclosed are methods utilizing the device.
Type:
Grant
Filed:
January 9, 2007
Date of Patent:
November 22, 2011
Assignee:
Applied Biosystems, LLC
Inventors:
Timothy Woudenberg, Michael Albin, Reid B. Kowallis, Yefim Raysberg, Robert P. Ragusa, Emily S. Winn-Deen
Abstract: The present invention concerns an isolated siRNA of from about 5 to about 20 nucleotides that mediates RNA interference. Also disclosed are methods of reducing expression of a target gene in a cell comprising obtaining at least one siRNA of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 basepairs in length; and delivering the siRNA into the cell. The siRNAs can be chemically synthesized RNA or an analog of a naturally occurring RNA.
Abstract: The present invention encompasses methods for producing a modified polynucleotide sequence that comprises a (e.g., one or more) phosphorothiolate linkage, methods for determining a polynucleotide sequence comprising a (e.g., one or more) phosphorothiolate linkage, and methods for separating forward and reverse extension products that comprise a (e.g., one or more) phosphorothiolate linkage. The invention also encompasses kits for producing and/or determining the sequence of a modified polynucleotide that comprises a (e.g., one or more) phosphorothiolate linkage.