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    DNA sequencing and DNA terminators
    1.
    发明授权
    DNA sequencing and DNA terminators 失效
    DNA测序和DNA终止子

    公开(公告)号：US06207421B1

    公开(公告)日：2001-03-27

    申请号：US08505072

    申请日：1995-07-21

    申请人： Lyle Richard Middendorf ,  John A. Brumbaugh

    发明人： Lyle Richard Middendorf ,  John A. Brumbaugh

    IPC分类号： C12Q168

    CPC分类号： G01N27/44721 ,  C07D209/60 ,  C09B23/0066 ,  C12Q1/6869 ,  G01N21/6428 ,  G01N27/447 ,  G01N27/44726 ,  C12Q2525/186 ,  C12Q2537/161 ,  C12Q2535/101 ,  C12Q2525/101

    摘要： A universal terminator includes a heterocycle other than naturally occuring DNA heterocycles such as adenine, cytosine, guanine and thymine. In terminating strand synthesis, the hybridized primer-template is split into four aliquots. In the “A” vial is added DNA polymerase and normal amounts of “C”, “G”, and “T” deoxynucleotides, along with a reduced amount of “A” deoxynucleotide and an amount of the universal terminator such that statistically the universal terminator has less than a one percent chance of being incorporated at sites where a “C”, “G”, or “T” deoxynucleotide should be incorporated and about a one percent chance of being incorporated at sites where an “A” deoxynucleotide should be incorporated. Similar strategies are followed for the “C”, “G”, and “T” vials, except that the amount of “C”, “G” and “T” deoxynucleotides are reduced for their respective vial.

    摘要翻译： 通用终止子包括除天然存在的杂环之外的杂环，例如腺嘌呤，胞嘧啶，鸟嘌呤和胸腺嘧啶。 在末端链合成中，将杂交的引物 - 模板分成四个等分试样。 在“A”小瓶中加入DNA聚合酶和正常量的“C”，“G”和“T”脱氧核苷酸，以及减少量的“A”脱氧核苷酸和通用终止子的量，使得统计学上普遍 终止子具有少于1％的可能性，其中应加入“C”，“G”或“T”脱氧核苷酸的位点，并且在“A”脱氧核苷酸应位于“A” 并入。 对于“C”，“G”和“T”小瓶，遵循类似的策略，不同之处在于它们各自的小瓶中的“C”，“G”和“T”脱氧核苷酸的量被减少。

    DNA sequencing
    3.
    发明授权
    DNA sequencing 失效
    DNA测序

    公开(公告)号：US4729947A

    公开(公告)日：1988-03-08

    申请号：US594676

    申请日：1984-03-29

    申请人： Lyle R. Middendorf ,  John A. Brumbaugh

    发明人： Lyle R. Middendorf ,  John A. Brumbaugh

    IPC分类号： C07D209/60 ,  C12N15/00 ,  C12N15/09 ,  C12Q1/68 ,  G01N21/64 ,  G01N27/26 ,  G01N27/447 ,  G01N33/50 ,  G01N33/58

    CPC分类号： G01N27/44726 ,  C07D209/60 ,  C12Q1/6816 ,  C12Q1/6869 ,  G01N21/6428 ,  G01N27/447 ,  G01N27/44721 ,  Y10S435/803

    摘要： To sequence long strands of DNA, cloned strands having lengths longer than 100 bases are, in one embodiment, marked on one end with biotin. These strands are divided into 4 aliquots and each aliquot: (1) is uniquely chemically treated to randomly terminate the strands at the non-biotinylated end at a selected type of base; and (2) is moved continuously by electrophoresis through a different one of four identical channels. In the one embodiment, the strands are randomly terminated at a selected base type and they are moved into avidin, which due to high affinity, combines with the biotin marked ends of shorter strands before the longer strands are fully resolved in the gel. The avidin is marked with fluorescein, the strands are scanned and the signals are decoded. In another embodiment, the strands are synthesized, with termination at a selected base type and marked either by the above method or by ethidium bromide.

    摘要翻译： 为了序列长链DNA，在一个实施方案中，长度超过100个碱基的克隆的链在一端用生物素标记。 将这些链分成4等份，并且每个等分试样：（1）被唯一化学处理以在选定类型的碱基处的非生物素化末端随机终止链; 和（2）通过电泳通过四个相同通道中的不同的一个连续移动。 在一个实施方案中，链以选定的碱基类型随机终止，并且它们被移动到抗生物素蛋白中，其由于高亲和力而与长链在凝胶中完全分解之前与短链的生物素标记的末端结合。 抗生物蛋白标记有荧光素，扫描股线并对信号进行解码。 在另一个实施方案中，合成链，其终止于选定的碱基类型，并用上述方法或溴化乙锭标记。

    Digital DNA typing
    4.
    发明授权
    Digital DNA typing 失效

    公开(公告)号：US5549805A

    公开(公告)日：1996-08-27

    申请号：US288989

    申请日：1994-08-11

    申请人： Lyle R. Middendorf ,  John A. Brumbaugh ,  Gi Y. Jang

    发明人： Lyle R. Middendorf ,  John A. Brumbaugh ,  Gi Y. Jang

    IPC分类号： C07D209/60 ,  C12Q1/68 ,  G01N21/64 ,  G01N27/447 ,  G01N27/26

    CPC分类号： G01N21/6458 ,  C07D209/60 ,  C12Q1/6869 ,  G01N21/6428 ,  G01N27/447 ,  G01N27/44721 ,  G01N27/44726

    摘要： To sequence DNA automatically, fluorescently marked DNA are electrophoresed in a plurality of channels through a gel electrophoresis slab; wherein the DNA samples are resolved in accordance with the size of DNA fragments in the gel electrophoresis slab into fluorescently marked DNA bands. The separated samples are scanned photoelectrically with a laser and a sensor, wherein the laser scans with scanning light at a scanning light frequency within the absorbance spectrum of said fluorescently marked DNA samples and light is sensed at the emission frequency of the marked DNA. The light is modulated from said laser at a predetermined modulation frequency and fluorescent light emitted by said DNA bands at said modulation frequency is detected, whereby background noise from the medium through which the light is transmitted is discriminated against.

    DNA sequencing
    5.
    发明授权
    DNA sequencing 失效
    DNA测序

    公开(公告)号：US5346603A

    公开(公告)日：1994-09-13

    申请号：US950734

    申请日：1992-09-24

    申请人： Lyle R. Middendorf ,  John A. Brumbaugh

    发明人： Lyle R. Middendorf ,  John A. Brumbaugh

    IPC分类号： C07D209/60 ,  C12Q1/68 ,  G01N21/64 ,  G01N27/447 ,  C25B9/00

    CPC分类号： G01N21/6452 ,  C07D209/60 ,  C12Q1/6869 ,  G01N21/6428 ,  G01N27/447 ,  G01N27/44721 ,  G01N27/44726

    摘要： To sequence long strands of DNA, cloned strands having lengths longer than 100 bases are, in one embodiment, marked on one end with biotih. These strands are divided into 4 aliquots and each aliquot: (1) is uniquely chemically treated to randomly terminate the strands at the non-biotinylated end at a selected type of base; and (2) is moved continuously by electrophoresis through a different one of four identical channels. In the one embodiment, the strands are randomly terminated at a selected base type and they are moved into avidin, which due to high affinity, combines with the biotin marked ends of shorter strands before the longer strands are fully resolved in the gel. The avidin is marked with fluorescein, the strands are scanned and the signals are decoded. In another embodiment, the strands are synthesized, with termination at a selected base type and marked either by the above method or by ethidium bromide.

    摘要翻译： 为了排列长链DNA，在一个实施方案中，长度超过100个碱基的克隆的链在一端用生物标记。 将这些链分成4等份，并且每个等分试样：（1）被唯一化学处理以在选定类型的碱基处的非生物素化末端随机终止链; 和（2）通过电泳通过四个相同通道中的不同的一个连续移动。 在一个实施方案中，链以选定的碱基类型随机终止，并且它们被移动到抗生物素蛋白中，其由于高亲和力而与长链在凝胶中完全分解之前与短链的生物素标记的末端结合。 抗生物蛋白标记有荧光素，扫描股线并对信号进行解码。 在另一个实施方案中，合成链，其终止于选定的碱基类型，并用上述方法或溴化乙锭标记。

    DNA sequencing
    6.
    发明授权
    DNA sequencing 失效
    DNA测序

    公开(公告)号：US5207880A

    公开(公告)日：1993-05-04

    申请号：US570503

    申请日：1990-08-21

    申请人： Lyle R. Middendorf ,  John A. Brumbaugh

    发明人： Lyle R. Middendorf ,  John A. Brumbaugh

    IPC分类号： C12M1/00 ,  C07D209/60 ,  C12N15/09 ,  C12Q1/68 ,  G01N21/64 ,  G01N27/447 ,  G01N33/53

    CPC分类号： G01N27/447 ,  C07D209/60 ,  C12Q1/6869 ,  G01N21/6428 ,  G01N27/44721 ,  G01N27/44726 ,  G01N21/6458

    摘要： To sequence DNA automatically, flourescently marked DNA are electrophoresed in a plurality of channels through a gel electrophoresis slab; wherein the DNA samples are resolved in accordance with the size of DNA fragments in the gel electrophoresis slab into fluorescently marked DNA bands. The separated samples are scanned photoelectrically with a laser and a sensor, wherein the laser scans with scanning light at a scanning light frequency within the absorbance spectrum of said fluorescently marked DNA samples and light is sensed at the emission frequency of the marked DNA. The light is modulated from said laser at a predetermined modulation frequency and fluorescent light emitted by said DNA bands at said modulation frequency is detected, whereby background noise from the medium through which the light is transmitted is discriminated against.

    摘要翻译： 为了自动序列DNA，荧光标记的DNA通过凝胶电泳板在多个通道中进行电泳; 其中根据凝胶电泳板中DNA片段的大小将DNA样品分解成荧光标记的DNA条带。 分离的样品用激光和传感器进行光电扫描，其中激光以扫描光频率在所述荧光标记的DNA样品的吸收光谱内扫描，并以标记的DNA的发射频率感测光。 以预定的调制频率从所述激光器调制光，并且检测由所述DNA条带以所述调制频率发射的荧光，从而区分来自发射光的介质的背景噪声。