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1. 发明申请

WO1998008978A1 APPARATUS AND METHOD FOR PERFORMING SEQUENCING OF NUCLEIC ACID POLYMERS 审中-公开
标题翻译：用于实施核酸聚合物测序的装置和方法
公开(公告)号：WO1998008978A1
公开(公告)日：1998-03-05
申请号：PCT/US1997015056
申请日：1997-08-27
申请人： VISIBLE GENETICS INC. , WATERHOUSE, Paul , IZMAILOV, Alexandre, M. , ZALESKI, Henryk , YAGER, Thomas, D. , DUNN, James, M. , LEUSHNER, James , HUI, May , LARSON, Marina, T.
发明人： VISIBLE GENETICS INC.
IPC分类号： C12Q01/68
CPC分类号： B01L7/52 , G01N27/44743
摘要： An apparatus for processing samples containing DNA to produce a sequencing fragment mixture comprises a sample processing element comprising: a thermocycling region having one or more chambers for receiving a DNA sequencing reaction mixture and forming sequencing fragments therefrom; a separation region comprising a separation matrix for separating the sequencing fragments formed in the thermocycling regions; a detection region for detection of the separated sequencing fragments; and means for regulating the temperature in the thermocycling region of the sample processing element to provide a plurality of thermal cycles, each cycle including at least a denaturation phase and an extension phase. The apparatus for processing sample can be placed in a holder which is associated with means for applying an electric field to the separation region of a sample processing apparatus placed within the holder to cause polynucleotide sequencing fragments to migrate through the separation region from the thermocycling region to the detection region; and means for detecting polynucleotide fragments within the detection region of the sample processing apparatus placed within the holder.
摘要翻译：用于处理含有DNA的样品以产生测序片段混合物的装置包括样品处理元件，其包括：热循环区域，其具有用于接收DNA测序反应混合物并形成测序片段的一个或多个室; 分离区域，其包含用于分离在所述热循环区域中形成的测序片段的分离基质; 用于检测分离的测序片段的检测区域; 以及用于调节样品处理元件的热循环区域中的温度以提供多个热循环的装置，每个循环至少包括变性相和延伸相。用于处理样品的装置可以放置在与将电场施加到放置在保持器内的样品处理装置的分离区域的装置相关联的保持器中，以使得多核苷酸测序片段通过分离区域从热循环区域迁移到检测区域; 以及用于检测放置在保持器内的样品处理装置的检测区域内的多核苷酸片段的装置。

2. 发明申请

WO1996042012A1 NANOFABRICATED SEPARATION MATRIX FOR ANALYSIS OF BIOPOLYMERS AND METHODS OF MAKING AND USING SAME 审中-公开
标题翻译：用于生物聚合物分析的纳米分离基质及其制备方法和使用方法
公开(公告)号：WO1996042012A1
公开(公告)日：1996-12-27
申请号：PCT/US1996009999
申请日：1996-06-07
申请人： VISIBLE GENETICS INC. , YAGER, Thomas, D. , WATERHOUSE, Paul , IZMAILOV, Alexandre M. , MARUZZO, Bruno , STEVENS, John, K. , LARSON, Marina, T.
发明人： VISIBLE GENETICS INC.
IPC分类号： G01N27/447
CPC分类号： G01N27/44773 , G01N27/44704 , G01N27/44791
摘要： Separation matrices useful in the formation of solid-state mm- to cm-scale devices for the rapid, high-resolution separation of single-stranded DNA ladder bands generated by the Sanger dideoxy- or Maxam/Gilbert chemical DNA sequencing procedures are formed from a solid support (1) having a plurality of posts (4) disposed on a first major surface thereof to form an obstacle course of posts (4) and pores (5). The posts are arranged in a regular X, Y array and are separated one from another by a distance of 100 nm or less, preferably 10 to 30 nm, and are optionally separated into lanes 2. The separation matrix can be manufactured by first forming a mold, preferably a reusable mold using lithography techniques. The mold is the reverse of the desired pattern of posts and pores of the obstacle course, and is used for casting the obstacle course. The cast obstacle course is then fused to a solid support and separated from the mold. Alternatively, the separation matrix can be formed from a polymer which undergoes specific and quantifiable swelling in the presence of a selected chemical compound. In this case, the matrix is cast on a mold in a conventional manner with a spacing between posts greater than the desired final spacing of 100 nm or less. For use, a buffer solution saturated with the specific chemical agent that controls swelling is added, causing the posts to swell to a defined amount to achieve the desired separation.
摘要翻译：用于形成用于通过Sanger双脱氧或Maxam / Gilbert化学DNA测序程序产生的单链DNA梯形带的快速，高分辨率分离的固态mm至cm尺度装置的分离基质由固体支撑件（1）具有设置在其第一主表面上的多个柱（4），以形成柱（4）和孔（5）的障碍物路线。柱以规则的X，Y阵列布置，并且彼此分离一个距离为100nm或更小，优选为10至30nm，并且任选地分离成通道2.分离基体可以通过首先形成模具，优选使用光刻技术的可重复使用的模具。模具与障碍物路线的柱和孔的期望图案相反，并且用于铸造障碍物路线。然后将铸造的障碍物道路融合到固体支撑物并与模具分离。或者，分离基质可以由在所选择的化合物存在下经历特异性和可定量溶胀的聚合物形成。在这种情况下，基体以常规方式在模具上铸造，柱之间的间距大于期望的最终间距为100nm或更小。为了使用，加入饱和了特定化学试剂的缓冲溶液以控制溶胀，导致柱膨胀至规定的量以达到所需的分离。

3. 发明申请

WO9708344A2 METHOD FOR IDENTIFICATION OF MUTATIONS USING LIGATION OF MULTIPLE OLIGONUCLEOTIDE PROBES 审中-公开
标题翻译：使用多个寡核苷酸探针识别突变体的方法
公开(公告)号：WO9708344A2
公开(公告)日：1997-03-06
申请号：PCT/US9614020
申请日：1996-08-28
申请人： VISIBLE GENETICS INC , YAGER THOMAS D , DUNN JAMES M
发明人： YAGER THOMAS D , DUNN JAMES M
IPC分类号： C12N15/09 , C12Q1/68
CPC分类号： C12Q1/6827 , C12Q2563/107 , C12Q2533/107 , C12Q2527/107
摘要： A ligase-based assay which relies only upon knowledge of the wild-type sequence of a gene or gene fragment is used to detect all types of mutations, i.e., point mutations, insertions and deletions. The assay makes use of a set of oligonucleotide probes, which may be packaged in kit form, which hybridize in series along the length of the gene. The ligation of the probes together form a ligation product, the size of which is evaluated. When the gene or gene fragment being analyzed corresponds to the normal sequence and thus perfectly matches the probes, all of the probes in the set are ligated together, and the ligation product has a certain resulting size. When a mutation appears in the gene, the hybridization of the probe overlapping the mutation is impaired, with the result that some or all of the ligation product is of smaller size. By evaluating the size of the ligation product, both the existence of a mutation and its approximate position can be identified.
摘要翻译：仅依赖于基因或基因片段的野生型序列的知识的基于连接酶的测定法用于检测所有类型的突变，即点突变，插入和缺失。该测定使用一组寡核苷酸探针，其可以以试剂盒形式包装，其沿着该基因的长度串联杂交。探针的连接形成连接产物，评价其尺寸。当分析的基因或基因片段对应于正常序列，从而完美匹配探针时，将所有组中的所有探针连接在一起，并且连接产物具有一定的最终尺寸。当基因中出现突变时，与突变重叠的探针的杂交受损，结果是部分或全部连接产物的尺寸较小。通过评估连接产物的大小，可以确定突变的存在及其近似位置。

4. 发明申请

WO1997008344A3 METHOD FOR IDENTIFICATION OF MUTATIONS USING LIGATION OF MULTIPLE OLIGONUCLEOTIDE PROBES 审中-公开
公开(公告)号：WO1997008344A3
公开(公告)日：1997-03-06
申请号：PCT/US1996014020
申请日：1996-08-28
申请人： VISIBLE GENETICS INC. , YAGER, Thomas, D. , DUNN, James, M.
发明人： VISIBLE GENETICS INC.
IPC分类号： C12Q01/68

5. 发明申请

WO1996042013A1 MICROELECTROPHORESIS CHIP FOR MOVING AND SEPARATING NUCLEIC ACIDS AND OTHER CHARGED MOLECULES 审中-公开
标题翻译：用于移动和分离核酸和其他充电分子的微电极芯片
公开(公告)号：WO1996042013A1
公开(公告)日：1996-12-27
申请号：PCT/US1996010110
申请日：1996-06-07
申请人： VISIBLE GENETICS INC. , YAGER, Thomas, D. , WATERHOUSE, Paul , IZMAILOV, Alexandre, M. , MARUZZO, Bruno , STEVENS, John, K. , LARSON, Marina, T.
发明人： VISIBLE GENETICS INC.
IPC分类号： G01N27/447
CPC分类号： G01N27/44773 , G01N27/44704 , G01N27/44791
摘要： A microelectrophoresis chip comprises a substrate in which there are formed one or more channels, one channel for each sample to be evaluated. The channels extend for the length of the chip, a distance of generally around 1 cm, and are about 1 to 10 mu m wide and 1 to 10 mu m in depth. The channels are filled with a homogeneous separation matrix which acts as an obstacle to the electrophoretic migration of the charged molecules. Microelectrodes disposed in the channels are used to induce an electric field within the homogeneous separation medium. When a voltage is applied across two or more of the microelectrodes, the charged molecules are induced to move and separate according to the electric field density, the type of solvent film, and the charge, shape and size of the charged molecule. The chip may further comprise detectors, such as light polarization detectors, fluorescence emission detectors, biosensors, electrochemical sensors or other microcomponents which may include sites for enzymatic or chemical manipulation of the moved or separated charged molecules.
摘要翻译：微电泳芯片包括其中形成一个或多个通道的基底，每个待评估样品的通道一个通道。通道延长芯片的长度，通常为1cm左右，宽度为1〜10μm左右，深度为1〜10μm。通道充满均匀的分离基质，其作为电荷分子的电泳迁移的障碍。设置在通道中的微电极用于在均匀分离介质内诱发电场。当电压施加到两个或更多个微电极上时，根据电场密度，溶剂膜的类型和带电分子的电荷，形状和尺寸，诱导带电分子移动和分离。芯片还可以包括诸如光偏振检测器，荧光发射检测器，生物传感器，电化学传感器或其它微元件的检测器，其可以包括用于酶或分离带电分子的酶或化学操作的位点。

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