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1. 发明授权

US07655407B2 Method for detecting target substance utilizing probe desorption 有权
标题翻译：利用探针解吸法检测目标物质的方法
公开(公告)号：US07655407B2
公开(公告)日：2010-02-02
申请号：US12029840
申请日：2008-02-12
申请人： Hayato Miyoshi , Takeshi Senga
发明人： Hayato Miyoshi , Takeshi Senga
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6816 , C12Q2563/155 , C12Q2563/149 , C12Q2563/143
摘要： It is an object of the present invention to provide a method for detecting a target substance in a specimen with the use of fine particles, whereby the target substance can be readily detected with the exclusive use of a single type of probe and the detection limit is improved. The present invention provides a method for detecting a target substance in a specimen which comprises the steps of: allowing a complex of a fine particle and a probe to come into contact with a specimen; and detecting changes in physical properties of the fine particle that are caused by desorption of the probe from the fine particle due to interaction between the target substance in the specimen and the probe.
摘要翻译：本发明的目的是提供一种使用细小颗粒检测样品中的目标物质的方法，由此可以通过使用单一探针来容易地检测目标物质，检测限为改进。本发明提供一种检测试样中的目标物质的方法，包括以下步骤：使微粒和探针的复合物与样品接触; 并且由于样品中的目标物质与探针之间的相互作用，检测由微粒解吸所引起的微粒的物理性质的变化。

2. 发明授权

US08309305B2 Method for discriminating between nucleotide sequences of nucleic acids 有权
标题翻译：鉴别核酸核苷酸序列的方法
公开(公告)号：US08309305B2
公开(公告)日：2012-11-13
申请号：US12471817
申请日：2009-05-26
申请人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
发明人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6844 , C12Q1/6858 , C12Q2531/119 , C12Q2531/101
摘要： The present invention describes methods for discriminating between nucleotide sequences of first and second nucleic acids, including: providing a reaction solution, including a deoxynucleotide triphosphate, a DNA polymerase with strand displacement ability, a template nucleic acid fragment, a primer, and a mask oligo; incubating the reaction solution, obtaining a polymerase reaction, and producing an amplification product; detecting the amplification product to discriminate between the nucleotide sequences, wherein the primer is complementary to the first nucleic acid, the mask oligo hybridizes to the nucleotide sequence portion of the first and second nucleic acid, wherein the mask oligo is more complementary to the second nucleic acid than to the first nucleic acid, and wherein the mask oligo is not an origin of an elongation reaction with the polymerase, and a primer portion and a mask oligo portion hybridize to the same regions on the first and second nucleic acid.
摘要翻译：本发明描述了区分第一和第二核酸的核苷酸序列的方法，包括：提供包括脱氧核苷酸三磷酸的反应溶液，具有链置换能力的DNA聚合酶，模板核酸片段，引物和掩蔽寡核苷酸 ; 孵育反应溶液，获得聚合酶反应，并产生扩增产物; 检测扩增产物以区分其中引物与第一核酸互补的核苷酸序列，掩蔽寡核苷酸与第一和第二核酸的核苷酸序列部分杂交，其中掩蔽寡核苷酸与第二核酸更互补酸，而不是第一核酸，并且其中掩模寡核苷酸不是与聚合酶的延伸反应的起源，并且引物部分和掩模寡核苷酸部分与第一和第二核酸上的相同区域杂交。

3. 发明申请

US20090162856A1 RNA DETECTION METHOD 审中-公开
标题翻译： RNA检测方法
公开(公告)号：US20090162856A1
公开(公告)日：2009-06-25
申请号：US12265629
申请日：2008-11-05
申请人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
发明人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/686
摘要： It is an object of the present invention to provide a method for rapid, convenient, and highly sensitive detection of trace RNA wherein a risk of contamination is low. The present invention provides a method for amplification of nucleic acid which comprises the steps of: (i) allowing a reverse transcriptase to act on RNA so as to produce a nucleic acid fragment; and (ii) performing substantially isothermal incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase having strand displacement activity, a divalent cation, a surfactant accounting for at least 0.01% of the solution, at least two types of oligonucleotide primers, and the nucleic acid fragment as a template obtained in the step (i) so as to perform a polymerase reaction that is initiated from the 3′ ends of the primers and thus amplify the nucleic acid fragment.
摘要翻译：本发明的目的是提供一种用于快速，方便和高灵敏度地检测痕量RNA的方法，其中污染的风险低。本发明提供核酸扩增方法，其包括以下步骤：（i）使逆转录酶作用于RNA以产生核酸片段; 和（ii）对包含至少一种类型的脱氧核苷酸三磷酸的反应溶液，至少一种具有链置换活性的DNA聚合酶，二价阳离子，占该溶液的至少0.01％的表面活性剂进行基本上等温温育至少两种寡核苷酸引物，以及作为在步骤（i）中获得的模板的核酸片段，以进行从引物的3'末端引发的聚合酶反应，从而扩增核酸片段。

4. 发明授权

US07741441B2 Method for producing type IV collagen 有权
标题翻译： IV型胶原蛋白的制备方法
公开(公告)号：US07741441B2
公开(公告)日：2010-06-22
申请号：US11727573
申请日：2007-03-27
申请人： Takayuki Kobayashi , Tasuku Sasaki , Hayato Miyoshi
发明人： Takayuki Kobayashi , Tasuku Sasaki , Hayato Miyoshi
IPC分类号： C07K14/00
CPC分类号： C07K14/78
摘要： An object of the invention is to provide type IV collagen without contamination by other proteins and without degradation or denaturation. The present invention provides a type IV collagen which is derived and extracted from lens capsules without the use of an enzyme and has a minimum molecular weight of 160 to 180 kDa measured by SDS-PAGE under reduced conditions.
摘要翻译：本发明的目的是提供IV型胶原蛋白，而不会被其他蛋白质污染，而不会降解或变性。本发明提供IV型胶原蛋白，其在不使用酶的情况下从透镜胶囊衍生和提取，并且在减少的条件下通过SDS-PAGE测量的最小分子量为160至180kDa。

5. 发明申请

US20070232787A1 Method for producing type IV collagen 有权
标题翻译： IV型胶原蛋白的制备方法
公开(公告)号：US20070232787A1
公开(公告)日：2007-10-04
申请号：US11727573
申请日：2007-03-27
申请人： Takayuki Kobayashi , Tasuku Sasaki , Hayato Miyoshi
发明人： Takayuki Kobayashi , Tasuku Sasaki , Hayato Miyoshi
IPC分类号： C07K14/78
CPC分类号： C07K14/78
摘要： An object of the invention is to provide type IV collagen without contamination by other proteins and without degradation or denaturation. The present invention provides a type IV collagen which is derived and extracted from lens capsules without the use of an enzyme and has a minimum molecular weight of 160 to 180 kDa measured by SDS-PAGE under reduced conditions.
摘要翻译：本发明的目的是提供IV型胶原蛋白，而不会被其他蛋白质污染，而不会降解或变性。本发明提供IV型胶原蛋白，其在不使用酶的情况下从透镜胶囊衍生和提取，并且在减少的条件下通过SDS-PAGE测量的最小分子量为160至180kDa。

6. 发明申请

US20100047794A1 METHOD FOR DISCRIMINATING BETWEEN NUCLEOTIDE SEQUENCES OF NUCLEIC ACIDS 有权
标题翻译：用于分离核酸核苷酸序列的方法
公开(公告)号：US20100047794A1
公开(公告)日：2010-02-25
申请号：US12471817
申请日：2009-05-26
申请人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
发明人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6844 , C12Q1/6858 , C12Q2531/119 , C12Q2531/101
摘要： It is an object of the present invention to provide a method for discriminating between nucleic acid sequences with high accuracy by utilizing a method for specifically amplifying nucleic acid sequences under isothermal conditions. The present invention provides a method for discriminating between the nucleotide sequence of a first nucleic acid and the nucleotide sequence of a second nucleic acid by a nucleic acid amplification method that is performed under substantially isothermal conditions, wherein (1) at least one type of oligonucleotide (hereinafter “primer”) substantially complementary to the first nucleic acid, and (2) at least one type of oligonucleic acid (hereinafter “mask oligo”) that is designed such that it hybridizes to the nucleotide sequence portions of the first nucleic acid and the second nucleic acid to be discriminated, such that it is more complementary to the second nucleic acid than to the first nucleic acid, and such that it does not become an origin of an elongation reaction with polymerase, are used, the method being characterized in that a portion of the primer and a portion of the mask oligo hybridize to the same regions on the first nucleic acid and the second nucleic acid.
摘要翻译：本发明的目的是提供一种通过利用在等温条件下特异性扩增核酸序列的方法来高精度地区分核酸序列的方法。本发明提供了通过在基本上等温条件下进行的核酸扩增方法来区分第一核酸的核苷酸序列和第二核酸的核苷酸序列的方法，其中（1）至少一种类型的寡核苷酸（以下简称为“引物”），和（2）至少一种低聚核酸（以下称为“掩模寡核苷酸”），其被设计为与第一核酸的核苷酸序列部分杂交，要被鉴别的第二核酸，使得其比第一核酸与第二核酸更互补，并且使其不成为与聚合酶的延伸反应的起源，所述方法的特征在于引物的一部分和掩模寡核苷酸的一部分与第一核酸和第二核酸上的相同区域杂交。

7. 发明授权

US08435741B2 Isothermal nucleic acid amplification method using surfactant 有权
标题翻译：使用表面活性剂的等温核酸扩增方法
公开(公告)号：US08435741B2
公开(公告)日：2013-05-07
申请号：US12179098
申请日：2008-07-24
申请人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
发明人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6853 , C12Q2531/119 , C12Q2527/125 , C12Q2521/101
摘要： An object to be achieved by the present invention is to provide a nucleic acid amplification method by which a nucleic acid can be amplified substantially isothermally using oligonucleotide primers and DNA polymerase capable of strand displacement. The present invention provides a nucleic acid amplification method which comprises performing substantially isothermal incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase having strand displacement activity, a divalent cation, at least 0.01% or more surfactant, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3′ end of the primer and thus amplifying the nucleic acid fragment.
摘要翻译：本发明要实现的目的是提供一种使用寡核苷酸引物和能够进行链置换的DNA聚合酶基本上等温扩增核酸的核酸扩增方法。本发明提供了一种核酸扩增方法，其包括对含有至少一种类型的脱氧核苷酸三磷酸的反应溶液进行基本上恒温培养，至少一种具有链置换活性的DNA聚合酶，二价阳离子，至少0.01％或更多表面活性剂，至少两种类型的寡核苷酸引物和核酸片段作为模板，从而进行从引物的3'末端引发并因此扩增核酸片段的聚合酶反应。

8. 发明授权

US08420323B2 Nucleic acid amplification method 有权
标题翻译：核酸扩增法
公开(公告)号：US08420323B2
公开(公告)日：2013-04-16
申请号：US12270706
申请日：2008-11-13
申请人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
发明人： Hayato Miyoshi , Yoshihide Iwaki , Toshihiro Mori
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6844 , C12Q2531/119 , C12Q2525/155 , C12Q2525/161
摘要： An object to be achieved by the present invention is to provide a nucleic acid amplification method by which a nucleic acid can be amplified using oligonucleotide primers and DNA polymerase. The present invention provides a nucleic acid amplification method which comprises performing incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3′ end of the primer and thus amplifying the nucleic acid fragment, wherein a tag sequence is added at the 5′ end of the first oligonucleotide primer, and the tag sequence is a nucleotide sequence on the template nucleic acid fragment which is present downstream of the sequence which is substantially complementary with the 3′ end region of the first oligonucleotide primer (a region where the first oligonucleotide is annealed to the template nucleic acid).
摘要翻译：本发明要实现的目的是提供使用寡核苷酸引物和DNA聚合酶扩增核酸的核酸扩增方法。本发明提供核酸扩增方法，其包括对含有至少一种脱氧核苷酸三磷酸，至少一种DNA聚合酶，至少两种类型的寡核苷酸引物和核酸片段作为模板的反应溶液进行温育从而进行从引物的3'端引发的聚合酶反应，从而扩增核酸片段，其中在第一寡核苷酸引物的5'末端添加标签序列，并且标签序列是核苷酸序列在与第一寡核苷酸引物的3'末端区域（第一寡核苷酸与模板核酸退火的区域）基本上互补的序列下游的模板核酸片段上。

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