公开(公告)号：US20130344492A1

公开(公告)日：2013-12-26

申请号：US13916372

申请日：2013-06-12

申请人： Applied Biosystems, LLC

发明人： Peter MA

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6869 ,  C12Q2535/101 ,  C12Q2547/101 ,  C12Q2527/101

摘要： In one aspect, there are provided methods of amplifying and sequencing a polynucleotide. In some embodiments, the method includes (a) amplifying the polynucleotide with at least one amplification primer, a processive amplification polymerase, a sequencing primer, a sequencing polymerase, deoxynucleoside triphosphates suitable for template-dependent primer extension, and one or more terminating nucleotides, the incubation being carried out at a first temperature suitable for amplifying the polynucleotide with the processive amplification polymerase; (b) incubating the product of step (a) at a second temperature suitable for forming a plurality of differently-sized extended sequencing primers with the sequencing polymerase; (c) evaluating the extended sequencing primers in order to determine the sequence of the polynucleotide. The reactions at the first and second temperatures can be carried out in a single reaction vessel. In other aspects, compositions and kits for carrying out the methods are also provided.

摘要翻译： 一方面，提供了扩增和测序多核苷酸的方法。 在一些实施方案中，所述方法包括（a）用至少一个扩增引物，进程扩增聚合酶，测序引物，测序聚合酶，适于模板依赖性引物延伸的脱氧核苷三磷酸和一个或多个终止核苷酸扩增多核苷酸， 孵育在适于用进行性扩增聚合酶扩增多核苷酸的第一温度下进行; （b）在适于形成多个不同大小的扩增测序引物的第二温度下用所述测序聚合酶孵育步骤（a）的产物; （c）评估扩增的测序引物以确定多核苷酸的序列。 在第一和第二温度下的反应可以在单个反应容器中进行。 在其它方面，还提供了用于实施方法的组合物和试剂盒。

公开(公告)号：US20130261005A1

公开(公告)日：2013-10-03

申请号：US13905401

申请日：2013-05-30

申请人： APPLIED BIOSYSTEMS, LLC

发明人： Zheng ZHANG

IPC分类号： G06F19/22

CPC分类号： G16B30/00

摘要： Systems, methods, and analytical approaches for short read sequence assembly and for the detection of insertions and deletions (indels) in a reference genome. A method suitable for software implementation is presented in which indels may be readily identified in a computationally efficient manner.

摘要翻译： 用于短读序列组装和检测参考基因组中插入和缺失（indel）的系统，方法和分析方法。 提出了适用于软件实现的方法，其中可以以计算上有效的方式容易地识别输入。

公开(公告)号：US20130230915A1

公开(公告)日：2013-09-05

申请号：US13874110

申请日：2013-04-30

申请人： APPLIED BIOSYSTEMS, LLC

发明人： Hon Siu SHIN ,  Hock Lai KHOO

IPC分类号： B01L7/00

CPC分类号： B01L7/52 ,  B01L2300/0829 ,  B01L2300/1822 ,  B01L2300/1827

摘要： An apparatus and method for thermal cycling including a pasting edge heater. The pasting edge heater can provide substantial temperature uniformity throughout the retaining elements during thermal cycling by a thermoelectric module.

摘要翻译： 一种用于热循环的装置和方法，包括粘贴边缘加热器。 粘合边缘加热器可以通过热电模块在热循环期间在整个保持元件上提供显着的温度均匀性。

公开(公告)号：US20130177905A1

公开(公告)日：2013-07-11

申请号：US13650853

申请日：2012-10-12

申请人： Applied Biosystems, LLC

发明人： Richard A. Fekete ,  Annalee Nguyen

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6841 ,  C12N15/1003 ,  C12Q1/6844 ,  C12Q1/686

摘要： Sample preparation processes for in situ RNA or DNA analysis, methods and compositions therefor are provided. Processes provided herein allow DNA or RNA analysis to be carried out in the same tube or on an aliquot of the prepared sample without centrifugation or extraction. The preparation process can be carried out at room temperature in as little as seven minutes and is amenable to high throughput processing using manual or robotic platforms.

摘要翻译： 提供了用于原位RNA或DNA分析的样品制备方法，其方法和组合物。 本文提供的方法允许DNA或RNA分析在相同的管或制备样品的等分试样中进行，而无需离心或提取。 制备过程可以在室温下在短至七分钟内进行，并且可以使用手动或机器人平台进行高通量处理。

公开(公告)号：US20130115611A1

公开(公告)日：2013-05-09

申请号：US13673574

申请日：2012-11-09

申请人： Applied Biosystems, LLC

发明人： Stephen J. Gunstream

IPC分类号： G01N21/64

CPC分类号： C12Q1/6851 ,  G01N21/274 ,  G01N21/6428 ,  G01N21/6486 ,  G01N2021/6439 ,  G01N2021/6441 ,  G01N2201/12 ,  G01N2201/12746 ,  G16B30/00 ,  C12Q2545/113 ,  C12Q2545/101

摘要： The present teachings relate to a method of generating calibration information during a real-time polymerase chain reaction (RT-PCR) or other amplification reaction. A sample well plate or other support can contain one or more dyes or other reference materials that are subjected to the same RT-PCR thermal cycles or other conditions used to conduct amplification or other reactions on a biological sample. A set of maxima values and a set of minimum values, and/or other calibration information useful for adjusting emission data for sample dyes can be recorded, for example, for 10 cycles, 20 cycles, or each cycle of a complete RT-PCR run. Such testing of dye response under realistic operating conditions can enable more accurate characterization of plate, dye, filter, or instrument response and therefore more accurate calibration corrections and other and/or adjustments.

公开(公告)号：US11257570B2

公开(公告)日：2022-02-22

申请号：US15965380

申请日：2018-04-27

申请人： Applied Biosystems, LLC

发明人： Francis T. Cheng

IPC分类号： G16B40/10 ,  G16B45/00 ,  G16B99/00 ,  C12Q1/6844

摘要： Systems and methods are provided for processing a melting or dissociation curve of a DNA or other sample, for example, during PCR processing. In some embodiments, detection of the melting point and melting curve behavior can be enhanced by taking a derivative of the curve, and detecting peaks in the differential dissociation curve. In some embodiments, the derivative operation can comprise the use of edge-processing, or other detection algorithms. In some embodiments, the dissociation analysis can comprise removing low-frequency (or pedestal) components of the differential dissociation curve. In some embodiments, the differential dissociation curve can exhibit a smoothed or more regular appearance than the raw detected data.

公开(公告)号：US11225686B2

公开(公告)日：2022-01-18

申请号：US16746216

申请日：2020-01-17

申请人： APPLIED BIOSYSTEMS, LLC

发明人： Shoulian Dong ,  Junko F. Stevens ,  Danny H. Lee

IPC分类号： C12P19/34 ,  C12N15/115 ,  C12Q1/6844 ,  C12Q1/6848 ,  G01N21/64

摘要： The present teachings are directed to compositions, methods, and kits for amplifying target nucleic acids while reducing non-specific fluorescence and undesired amplification products, sometimes referred to as secondary amplification products or spurious side-products. The enzyme inhibitors disclosed herein comprise a nucleotide sequence and at least one quencher. Complexes comprising an enzyme inhibitor associated with an enzyme, wherein at least one enzymatic activity of the enzyme is inhibited, are also provided. Methods for amplifying a target nucleic acid while reducing undesired amplification products are disclosed, as are methods for reducing non-specific fluorescence. Kits for expediting the performance of certain disclosed methods are also provided.

公开(公告)号：US11175258B2

公开(公告)日：2021-11-16

申请号：US16247069

申请日：2019-01-14

申请人： APPLIED BIOSYSTEMS, LLC

发明人： Mark F. Oldham ,  Eric S. Nordman

IPC分类号： G01N27/414 ,  B82Y15/00 ,  C12Q1/6825 ,  G01N33/543

摘要： There is disclosed a system for electrical charge detection comprising a nanoFET device. Also disclosed is a method of electrical charge detection for single molecule sequencing. The method includes attaching a macromolecule or assemblies thereof to a gate of a nanoFET device and flowing in a solution of charge tags, where a charge tag includes a nucleotide attached to a charge complex. The method also includes incorporating one charge tag into the macromolecule or assemblies thereof and cleaving the charge tags from the macromolecule or assemblies thereof. The method further includes detecting at least one of current and voltage from the nanoFET device.

公开(公告)号：US11162137B2

公开(公告)日：2021-11-02

申请号：US16054391

申请日：2018-08-03

申请人： APPLIED BIOSYSTEMS, LLC

发明人： Linda G. Lee ,  Sam L. Woo ,  Congcong Ma ,  Richard T. Reel ,  Mark F. Oldham ,  David M. Cox ,  Benjamin G. Schroeder ,  Jon M. Sorenson ,  Willy Wiyatno

IPC分类号： C12Q1/6869 ,  B01L3/00 ,  F15C5/00 ,  F16K99/00 ,  G01N35/08 ,  G01N1/14 ,  C12Q1/6806 ,  C12Q1/6874 ,  G01N27/447 ,  B01L3/02 ,  B01L7/00

摘要： Various embodiments of the teachings relate to a system or method for sample preparation or analysis in biochemical or molecular biology procedures. The sample preparation can involve small volume processed in discrete portions or segments or slugs, herein referred to as discrete volumes. A molecular biology procedure can be nucleic acid analysis. Nucleic acid analysis can be an integrated DNA amplification/DNA sequencing procedure.

公开(公告)号：US11111525B2

公开(公告)日：2021-09-07

申请号：US15925119

申请日：2018-03-19

申请人： Applied Biosystems, LLC ,  AGILENT TECHNOLOGIES, INC.

发明人： Kirsten Vang Nielsen ,  Jens Hyldig-Nielsen ,  Brett F. Williams

IPC分类号： C07H21/04 ,  C12Q1/68 ,  C12Q1/6832 ,  C12Q1/6837 ,  C12Q1/6876

摘要： This invention is directed to methods, kits, non-nucleotide probes as well as other compositions pertaining to the suppression of binding of detectable nucleic acid probes to undesired nucleotide sequences of genomic nucleic acid in assays designed to determine target genomic nucleic acid.