公开(公告)号：US06534273B2

公开(公告)日：2003-03-18

申请号：US09956412

申请日：2001-09-18

申请人： William G. Weisburg ,  Jay H. Shaw ,  Michael M. Becker ,  Mehrdad R. Majlessi ,  Steven T. Brentano ,  Kiyotada Nunomura

发明人： William G. Weisburg ,  Jay H. Shaw ,  Michael M. Becker ,  Mehrdad R. Majlessi ,  Steven T. Brentano ,  Kiyotada Nunomura

IPC分类号： C12Q168

CPC分类号： C12Q1/6834 ,  C12Q1/6813

摘要： A method for capturing a target polynucleotide in a sample onto a solid support with an attached immobilized probe by using a capture probe and two different hybridization conditions that control the order of hybridization, where the first hybridization condition allows hybridization of the capture probe to the target polynucleotide, and the second hybridization condition allows hybridization of the capture probe to the immobilized probe. The method further includes amplifying the captured target polynucleotide by hybridizing at least one primer oligonucleotide to the target polynucleotide and using nucleic acid amplification that initiates from the primer oligonucleotide.

公开(公告)号：US5386024A

公开(公告)日：1995-01-31

申请号：US15729

申请日：1993-02-10

申请人： Daniel L. Kacian ,  Kiyotada Nunomura

发明人： Daniel L. Kacian ,  Kiyotada Nunomura

IPC分类号： C07H1/08 ,  C07H21/04 ,  C07K14/18 ,  C12N9/64 ,  C12N15/09 ,  C12N15/10 ,  C12Q1/68 ,  C07H23/00

CPC分类号： C07K14/005 ,  C12N15/1003 ,  C12N9/6481 ,  C12Q1/6806 ,  C12N2770/24222 ,  Y10S435/81 ,  Y10T436/25125

摘要： Method for making available a desired nucleic acid contained in a biological sample, comprising the steps of acidifying said biological sample to a pH at which endogenous nucleases capable of degrading the desired nucleic acid(s) are inactive, contacting said biological sample with an exogenous acid protease active at said pH, incubating said sample until endogenous nuclease activities are reduced to insignificant levels, and raising the pH of the biological sample to a pH sufficient to render the exogenous protease less active.

摘要翻译： 使生物样品中含有所需核酸的方法包括以下步骤：将所述生物样品酸化至能够使所需核酸降解的内源性核酸酶无活性的pH，使所述生物样品与外源性酸接触 蛋白酶在所述pH下活跃，孵育所述样品，直到内源核酸酶活性降低至微不足道的水平，并将生物样品的pH提高到足以使外源蛋白酶较少活性的pH。

公开(公告)号：US06294338B1

公开(公告)日：2001-09-25

申请号：US09620958

申请日：2000-07-21

申请人： Kiyotada Nunomura

发明人： Kiyotada Nunomura

IPC分类号： C12Q168

CPC分类号： C12Q1/6865 ,  C12Q1/6851 ,  C12Q2545/107

摘要： Methods useful for improving results obtained with enzyme-based polynucleotide amplification reactions. More particularly, the invented methods are useful for: (1) promoting amplification of template-specific products such that the amount of amplicon produced reflects the pre-amplification amount of analyte, even in reactions primed with low levels of analyte polynucleotide; (2) facilitating biological specimen processing such that the amount of amplicon produced in subsequent amplification reactions will be substantially independent of the efficiency of analyte polynucleotide isolation from the specimen; and (3) controlling the amount of analyte amplicon produced in the amplification reaction.

摘要翻译： 用于改善用酶基多核苷酸扩增反应获得的结果的方法。 更具体地，本发明的方法可用于：（1）促进模板特异性产物的扩增，使得所产生的扩增子的量反映了分析物的预扩增量，即使在用低水平的分析物多核苷酸引发的反应中; （2）促进生物样品处理，使得在随后的扩增反应中产生的扩增子的量将基本上独立于来自样品的分析物多核苷酸分离的效率; 和（3）控制在扩增反应中产生的分析物扩增子的量。

公开(公告)号：US06908735B2

公开(公告)日：2005-06-21

申请号：US09943286

申请日：2001-08-30

申请人： Kiyotada Nunomura

发明人： Kiyotada Nunomura

IPC分类号： G01N33/53 ,  C12N15/09 ,  C12Q1/68 ,  G01N21/78 ,  G01N33/566 ,  G01N33/58 ,  C07H21/02 ,  C07H21/04 ,  C12P19/34

CPC分类号： C12Q1/6865 ,  C12Q1/6851 ,  C12Q2545/107

摘要： Methods useful for improving results obtained with enzyme-based polynucleotide amplification reactions. More particularly, the invented methods are useful for: (1) promoting amplification of template-specific products such that the amount of amplicon produced reflects the pre-amplification amount of analyte, even in reactions primed with low levels of analyte polynucleotide; (2) facilitating biological specimen processing such that the amount of amplicon produced in subsequent amplification reactions will be substantially independent of the efficiency of analyte polynucleotide isolation from the specimen; and (3) controlling the amount of analyte amplicon produced in the amplification reaction.

摘要翻译： 用于改善用酶基多核苷酸扩增反应获得的结果的方法。 更具体地，本发明的方法可用于：（1）促进模板特异性产物的扩增，使得所产生的扩增子的量反映了分析物的预扩增量，即使在用低水平的分析物多核苷酸引发的反应中; （2）促进生物样品处理，使得在随后的扩增反应中产生的扩增子的量将基本上独立于来自样品的分析物多核苷酸分离的效率; 和（3）控制在扩增反应中产生的分析物扩增子的量。

公开(公告)号：US06610475B1

公开(公告)日：2003-08-26

申请号：US08347272

申请日：1994-11-30

申请人： Daniel L. Kacian ,  Kiyotada Nunomura

发明人： Daniel L. Kacian ,  Kiyotada Nunomura

IPC分类号： C12Q168

CPC分类号： C07K14/005 ,  C12N9/6481 ,  C12N15/1003 ,  C12N2770/24222 ,  C12Q1/6806 ,  Y10S435/81 ,  Y10T436/25125

摘要： The present invention features kits for making available a desired nucleic acid contained in a biological sample. The kits contain an acid for acidifying the biological sample to a pH at which endogenous nucleases capable of degrading desired nucleic acids are inactive and an acid protease able to digest cellular materials in the biological sample to release nucleic acid and to inactivate endogenous nucleases which may be present.

摘要翻译： 本发明提供用于提供生物样品中包含的所需核酸的试剂盒。 试剂盒含有用于将生物样品酸化至能够降解所需核酸的内源核酸酶无活性的酸，以及能够消化生物样品中的细胞物质以释放核酸并灭活内源核酸酶的酸， 当下。