-
公开(公告)号:US07951536B2
公开(公告)日:2011-05-31
申请号:US12279924
申请日:2007-02-20
申请人: Takeharu Nagai , Tomomi Tani , Ippei Kotera , Yohihiro Yoneda
发明人: Takeharu Nagai , Tomomi Tani , Ippei Kotera , Yohihiro Yoneda
CPC分类号: C12Q1/6869 , C12Q2563/107 , C12Q2521/313 , C12Q2521/501
摘要: The present invention provides a method for determining the base sequence of a DNA. According to the method for determining the base sequence of a DNA of the present invention, a probe is used, which is a probe having a protruding end and identification-labeled according to the species of the base at the protruding end, containing a recognition sequence of a class IIS restriction enzyme, to carry out simultaneously in a chain reaction, for a plurality of DNAs to be analyzed, ligation of the end base of a DNA to be analyzed and a probe and cleavage of the end base of the DNA to be analyzed, allowing the base sequence to be determined sequentially by a single molecule spectrofluorimetry method, such that an effective determination of the base sequence of a DNA becomes possible.
摘要翻译: 本发明提供了确定DNA碱基序列的方法。 根据本发明的DNA的碱基序列的测定方法,使用探针,该探针是具有突出端的探针,根据突起端的碱基的种类鉴定标记,其含有识别序列 的IIS类限制酶,在连锁反应中同时进行多个要分析的DNA,连接待分析的DNA的末端碱基和探针,并将DNA的末端碱基切割成 分析,通过单分子分光荧光法依次确定碱基序列,使得DNA的碱基序列的有效测定成为可能。
-
公开(公告)号:US20110244521A1
公开(公告)日:2011-10-06
申请号:US12674160
申请日:2008-08-12
申请人: Takeharu Nagai , Ippei Kotera
发明人: Takeharu Nagai , Ippei Kotera
IPC分类号: C12P19/34
摘要: The problem to be solved in the present invention is to provide a simplified and efficiently improved DNA recombination method.The above problem can be solved with the present method for preparing a recombinant DNA by inserting a DNA fragment of interest into a vector DNA, the method comprising the step of carrying out the following reactions at the same reacting location at the substantially simultaneouse time: (a) a reaction for simultaneously cleaving a site of the vector for inserting the fragment and a DNA containing the fragment in the presence of a restriction enzyme whose DNA recognition site and DNA cleavage site are discrete; and (b) a reaction for inserting the fragment into the vector in the presence of a DNA ligase.
摘要翻译: 本发明要解决的问题是提供简化且有效改进的DNA重组方法。 通过将目的DNA片段插入载体DNA中制备重组DNA的本发明的方法可以解决上述问题,该方法包括在大致同步的时间在相同的反应位置进行以下反应的步骤:( a)在DNA识别位点和DNA切割位点离散的限制性酶的存在下,同时切割用于插入片段的载体的位点和含有该片段的DNA的反应; 和(b)在DNA连接酶存在下将片段插入载体的反应。
-
公开(公告)号:US20100009354A1
公开(公告)日:2010-01-14
申请号:US12279924
申请日:2007-02-20
申请人: Takeharu Nagai , Tomomi Tani , Ippei Kotera , Yoshihiro Yoneda
发明人: Takeharu Nagai , Tomomi Tani , Ippei Kotera , Yoshihiro Yoneda
CPC分类号: C12Q1/6869 , C12Q2563/107 , C12Q2521/313 , C12Q2521/501
摘要: The present invention provides a method for determining the base sequence of a DNA. According to the method for determining the base sequence of a DNA of the present invention, a probe is used, which is a probe having a protruding end and identification-labeled according to the species of the base at the protruding end, containing a recognition sequence of a class IIS restriction enzyme, to carry out simultaneously in a chain reaction, for a plurality of DNAs to be analyzed, ligation of the end base of a DNA to be analyzed and a probe and cleavage of the end base of the DNA to be analyzed, allowing the base sequence to be determined sequentially by a single molecule spectrofluorimetry method, such that an effective determination of the base sequence of a DNA becomes possible.
摘要翻译: 本发明提供了确定DNA碱基序列的方法。 根据本发明的DNA的碱基序列的测定方法,使用探针,该探针是具有突出端的探针,根据突起端的碱基的种类鉴定标记,其含有识别序列 的IIS类限制酶,在连锁反应中同时进行多个要分析的DNA,连接待分析的DNA的末端碱基和探针,并将DNA的末端碱基切割成 分析,通过单分子分光荧光法依次确定碱基序列,使得DNA的碱基序列的有效测定成为可能。
-
-