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公开(公告)号:US06200748B1
公开(公告)日:2001-03-13
申请号:US08484340
申请日:1995-06-07
申请人: Lloyd M. Smith , Leroy E. Hood , Michael W. Hunkapiller , Tim J. Hunkapiller , Charles R. Connell
发明人: Lloyd M. Smith , Leroy E. Hood , Michael W. Hunkapiller , Tim J. Hunkapiller , Charles R. Connell
IPC分类号: C12Q168
CPC分类号: C12Q1/6869 , C12Q1/6816 , G01N27/44721 , G01N27/44726 , C12Q2563/107 , C12Q2535/101
摘要: This invention provides a duplex comprising an oligonucleotide primer and a template, wherein the primer is coupled chemically to a chromophore or fluorophore so as to allow chain extension by a polymerase. In one embodiment, the primer is extended by a polymerase to generate the complement of the template. In a further embodiment, the extended primer is separated from the template for use in a number of methods, including sequencing reactions. Methods of generating these compositions of matter are further provided.
摘要翻译: 本发明提供了一种双链体,其包含寡核苷酸引物和模板,其中引物与发色团或荧光团化学偶联,以允许通过聚合酶进行链延伸。 在一个实施方案中,引物通过聚合酶延伸以产生模板的补体。 在另一个实施方案中,将延伸的引物与模板分离以用于多种方法,包括测序反应。 进一步提供产生这些物质组合物的方法。
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公开(公告)号:US5780233A
公开(公告)日:1998-07-14
申请号:US659605
申请日:1996-06-06
申请人: Zhen Guo , Lloyd M. Smith
发明人: Zhen Guo , Lloyd M. Smith
CPC分类号: C12Q1/6827
摘要: An improved nucleic acid hybridization process is provided which employs a modified oligonucleotide and improves the ability to discriminate a control nucleic acid target from a variant nucleic acid target containing a sequence variation. The modified probe contains at least one artificial mismatch relative to the control nucleic acid target in addition to any mismatch(es) arising from the sequence variation. The invention has direct and advantageous application to numerous existing hybridization methods, including, applications that employ, for example, the Polymerase Chain Reaction, allele-specific nucleic acid sequencing methods, and diagnostic hybridization methods.
摘要翻译: 提供了改进的核酸杂交方法,其使用经修饰的寡核苷酸,并提高了鉴别来自含有序列变异的变体核酸靶的对照核酸靶的能力。 修饰的探针除了由序列变异产生的任何不匹配之外,还包含相对于对照核酸靶的至少一个人工错配。 本发明对许多现有的杂交方法具有直接和有利的应用,包括使用例如聚合酶链式反应,等位基因特异性核酸测序方法和诊断杂交方法的应用。
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公开(公告)号:US5401632A
公开(公告)日:1995-03-28
申请号:US915245
申请日:1992-07-16
申请人: Renfeng Wang , Lloyd M. Smith , Xinchun E. Tong
发明人: Renfeng Wang , Lloyd M. Smith , Xinchun E. Tong
CPC分类号: C12Q1/6813 , C12N15/1006 , C12Q1/6869
摘要: Disclosed herein are methods, kits, and equipment for purifying single stranded circular DNA and then using the DNA for DNA sequencing purposes. Templates are provided with an insert having a hybridization region. An elongated oligonucleotide has two regions that are complementary to the insert and the oligo is bound to a magnetic anchor. The oligo hybridizes to the insert on two sides to form a stable triple helix complex. The anchor can then be used to drag the template out of solution using a magnet. The system can purify sequencing templates, and if desired the triple helix complex can be opened up to a double helix so that the oligonucleotide will act as a primer for further DNA synthesis.
摘要翻译: 本文公开了用于纯化单链环状DNA然后使用DNA进行DNA测序目的的方法,试剂盒和设备。 提供了具有杂交区域的插入物的模板。 细长的寡核苷酸具有与插入物互补的两个区域,寡聚体结合到磁性锚定体上。 寡核苷酸与两侧的插入物杂交以形成稳定的三螺旋复合物。 然后可以使用锚来使用磁体将模板从溶液中拖出。 该系统可以纯化测序模板,如果需要,三螺旋复合物可以打开至双螺旋,使得寡核苷酸将作为进一步DNA合成的引物。
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公开(公告)号:US5212304A
公开(公告)日:1993-05-18
申请号:US216768
申请日:1988-07-08
申请人: Steven Fung , Sam L. Woo , Lloyd M. Smith
发明人: Steven Fung , Sam L. Woo , Lloyd M. Smith
IPC分类号: C07H1/00 , C07F9/09 , C07F9/24 , C07F9/572 , C07F9/59 , C07F9/6509 , C07F9/6584 , C07H19/10 , C07H19/20 , C07H21/00 , C08B37/00 , C08B37/02
CPC分类号: C07F9/2416 , A61K47/48061 , A61K47/48084 , C07F9/2408 , C07F9/65842 , C07H21/00 , C08B37/0021 , C08B37/0039
摘要: The compounds of the invention include novel linking agents comprising 2-substituted-3-protected-1,3,2-oxazaphosphacycloalkanes and their phosphoramidite precursors. The compounds of the invention further include conjugates of the above linking agents with oligonucleotides and polymer supports. The compounds of the present invention are useful for linking organic moieties, such as fluorescent or chromogenic dyes, to polymer supports and oligonucleotides, particularly single- and double-stranded DNA and RNA fragments.
摘要翻译: 本发明的化合物包括包含2-取代-3-保护的1,3,2-氧杂氮杂环烷烃及其亚磷酰胺前体的新型交联剂。 本发明的化合物还包括上述连接剂与寡核苷酸和聚合物载体的缀合物。 本发明的化合物可用于将有机部分如荧光或显色染料与聚合物载体和寡核苷酸,特别是单链和双链DNA和RNA片段连接。
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