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11. 发明授权

US6033880A Nucleic acid amplification reaction apparatus and method 失效
标题翻译：核酸扩增反应装置及方法
公开(公告)号：US6033880A
公开(公告)日：2000-03-07
申请号：US27989
申请日：1998-02-23
申请人： Lawrence A. Haff , Enrico Picozza , Will Bloch , Robert Ragusa , Joseph DiCesare , David Tracy , Paul Saviano , Timothy M. Woudenberg , Richard W. Noreiks
发明人： Lawrence A. Haff , Enrico Picozza , Will Bloch , Robert Ragusa , Joseph DiCesare , David Tracy , Paul Saviano , Timothy M. Woudenberg , Richard W. Noreiks
IPC分类号： C12N15/09 , B01J19/00 , B01L7/00 , C12M1/00 , C12Q1/68 , C40B40/06 , C40B60/14 , G01N35/00 , C12P19/34
CPC分类号： B01L7/525 , B01J19/0046 , B01L7/52 , B01L9/065 , B01J2219/00495 , B01J2219/00722 , B01L2300/1883 , C12Q1/686 , C40B40/06 , C40B60/14 , G01N2035/00237 , Y10T436/116664
摘要： Apparatus and method for performing a nucleic acid amplification reaction and preferably a polymerase chain reaction (PCR) in a reaction mixture in at least one capillary tube. Several different embodiments are disclosed. One embodiment cycles a sample through a capillary tube loop passing through two thermostatted fluid baths. Another embodiment has the capillary tube routed alternatingly between two heat exchangers to that the sample makes only one pass through the tube. Other embodiments maintain the heat exchangers stationary and translate the samples between them. Still further embodiments maintain the samples stationary and either automatically translate or rotate the heat exchangers past the samples contained within the capillary tubes to perform the thermal cycles necessary for the amplification reaction.
摘要翻译：用于在至少一个毛细管中的反应混合物中进行核酸扩增反应和优选聚合酶链式反应（PCR）的装置和方法。公开了几个不同的实施例。一个实施例通过穿过两个恒温流体浴的毛细管回路循环样品。另一个实施例中，毛细管在两个热交换器之间交替布置，使得样品仅穿过管中一次。其他实施例保持热交换器平稳，并在它们之间平移样品。另外的实施例保持样品静止，并且自动地将热交换器平移或旋转通过毛细管内包含的样品，以进行扩增反应所需的热循环。

12. 发明授权

US5972618A Detection of mutations in nucleic acids by chemical cleavage 有权
标题翻译：通过化学裂解检测核酸中的突变
公开(公告)号：US5972618A
公开(公告)日：1999-10-26
申请号：US130258
申请日：1998-08-06
申请人： Will Bloch
发明人： Will Bloch
IPC分类号： G01N33/50 , C12N15/09 , C12Q1/68 , C07H21/02 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6827
摘要： Improved methods, compositions and kits for detecting mutations in nucleic acids are disclosed, such comprise annealing a piece of control nucleic acid without mutations to a piece of test nucleic acid very similar in sequence to the control nucleic acid but possibly containing mutations, treating this mixture with potassium permanganate or hydroxylamine to remove mismatched bases from the duplex nucleic acid, treating the resulting nucleic acid with any of a class of diamines, triamines, and tetraamines analogous to 1,2-ethylenediamine to cleave abasic sites, and then analyzing the chemically treated nucleic acid to determine whether cleavage has occurred and approximately at what position in the nucleic acid any cleavage has occurred. Inclusion of betaine in the base-removal reactions improves their sensitivity and specificity. This series of non-hazardous, non-noxious, chemical treatments can be performed rapidly in a single reaction tube with no or minimal intervening separation steps; and the cleavage product can be prepared for the analysis step without removing the chemical reagents used to effect cleavage.
摘要翻译：公开了改进的用于检测核酸突变的方法，组合物和试剂盒，这包括对一片对照核酸进行退火，而不突变到与对照核酸非常相似的测试核酸片段，但可能含有突变，处理该混合物用高锰酸钾或羟胺从双链核酸中除去错配的碱基，用类似于1,2-乙二胺的一类二胺，三胺和四胺处理所得核酸，以切割脱碱基位点，然后分析化学处理核酸，以确定是否已经发生切割，并且大约在核酸中发生任何切割的位置。在脱碱反应中包含甜菜碱可以提高其敏感性和特异性。这种无毒无毒化学处理可以在单个反应管中快速进行，没有或最小的介入分离步骤; 并且可以制备裂解产物用于分析步骤而不除去用于切割的化学试剂。

13. 发明授权

US5866429A Precision and accuracy of anion-exchange separation of nucleic acids 失效
公开(公告)号：US5866429A
公开(公告)日：1999-02-02
申请号：US515885
申请日：1995-08-16
申请人： Will Bloch
发明人： Will Bloch
IPC分类号： B01D15/36 , B01J41/20 , G01N27/447 , G01N30/02 , G01N30/34 , G01N30/88
CPC分类号： B01D15/363 , B01J41/20 , G01N30/34 , G01N2030/8813 , G01N30/02 , Y10T436/108331 , Y10T436/143333
摘要： Solvents for salt-gradient anion-exchange separation of nucleic acids, especially double-stranded DNA and especially by liquid chromatography, are improved by replacing NaCl as the eluting salt with any of a wide range of alkyl ammonium salts and can be used to elute nucleic acids in strict order of increasing length, with reduced sensitivity to elution temperature and salt concentration. Anion-exchange chromatography with these solvents is well suited for identification of DNA fragments on the basis of size, with greater accuracy, precision, and resolvable size range than often is possible with gel electrophoresis.

14. 发明授权

US5827480A Nucleic acid amplification reaction apparatus 失效
标题翻译：核酸扩增反应装置
公开(公告)号：US5827480A
公开(公告)日：1998-10-27
申请号：US823735
申请日：1997-03-25
申请人： Lawrence A. Haff , Enrico Picozza , Will Bloch , Timothy M. Woudenberg
发明人： Lawrence A. Haff , Enrico Picozza , Will Bloch , Timothy M. Woudenberg
IPC分类号： C12N15/09 , B01J19/00 , B01L7/00 , C12M1/00 , C12Q1/68 , C40B40/06 , C40B60/14 , G01N35/00 , G01N21/01 , C12M1/02 , G01N21/62 , G01N33/50
CPC分类号： B01L7/525 , B01J19/0046 , B01L7/52 , B01L9/065 , B01J2219/00495 , B01J2219/00722 , B01L2300/1883 , C12Q1/686 , C40B40/06 , C40B60/14 , G01N2035/00237 , Y10T436/116664
摘要： Apparatus and method for performing a nucleic acid amplification reaction and preferably a polymerase chain reaction (PCR) in a reaction mixture in at least one capillary tube. Several different embodiments are disclosed. One embodiment cycles a sample through a capillary tube loop passing through two thermostatted fluid baths. Another embodiment has the capillary tube routed alternatingly between two heat exchangers to that the sample makes only one pass through the tube. Other embodiments maintain the heat exchangers stationary and translate the samples between them. Still further embodiments maintain the samples stationary and either automatically translate or rotate the heat exchangers past the samples contained within the capillary tubes to perform the thermal cycles necessary for the amplification reaction.
摘要翻译：用于在至少一个毛细管中的反应混合物中进行核酸扩增反应和优选聚合酶链式反应（PCR）的装置和方法。公开了几个不同的实施例。一个实施例通过穿过两个恒温流体浴的毛细管回路循环样品。另一个实施例中，毛细管在两个热交换器之间交替布置，使得样品仅穿过管中一次。其他实施例保持热交换器平稳，并在它们之间平移样品。另外的实施例保持样品静止，并且自动地将热交换器平移或旋转通过毛细管内包含的样品，以进行扩增反应所需的热循环。

15. 发明授权

US4789630A Ionic compounds containing the cationic meriquinone of a benzidine 失效
标题翻译：含有联苯胺阳离子季戊四醇的离子化合物
公开(公告)号：US4789630A
公开(公告)日：1988-12-06
申请号：US896677
申请日：1986-08-20
申请人： Will Bloch , Patrick J. Sheridan , Robert J. Goodson
发明人： Will Bloch , Patrick J. Sheridan , Robert J. Goodson
IPC分类号： G01N33/545 , C12Q1/28 , C12Q1/68 , G01N21/82 , G01N33/535 , G01N33/543 , G01N33/58 , G01N33/53
CPC分类号： C12Q1/28 , C12Q1/68 , G01N33/581 , C12Q2326/12 , Y10S435/803 , Y10S435/81 , Y10S435/96 , Y10S435/975
摘要： Useful for visualizing biological materials in a solid phase, on a gel, or in a liquid phase is a solid salt of the meriquinone of benzidine or a substituted benzidine. An immobilized or dissolved complex of a polymeric anion and the meriquinone of benzidine or a substituted benzidine having controllable solubility may also be employed. Preferred are meriquinone salts and complexes of 3,3,5,5'-tetramethylbenzidine. For visualization, the benzidine or substituted benzidine is oxidized to its meriquinone at pH 3 to 7 in the presence of an effective anion or polymeric anion, an oxidation catalyst, and an effective amount of oxidant to form a solid salt or immobilized complex of the meriquinone under conditions where the meriquinone solubility lies below about 10.sup.-5 M.
摘要翻译：可用于在固相，凝胶或液相中可视化生物材料是联苯胺或取代的联苯胺的固醇的固体盐。还可以使用固定或溶解的聚合阴离子的复合物和具有可控的溶解度的联苯胺或取代的联苯胺的金刚酮。优选3,3,5,5'-四甲基联苯胺的季戊四醇盐和络合物。为了可视化，联苯胺或取代的联苯胺在有效阴离子或聚合阴离子，氧化催化剂和有效量的氧化剂的存在下，在pH 3至7下被氧化成其三聚物，以形成固体盐或固定的复合物在meriquinone溶解度低于约10 -5 M的条件下。

16. 发明申请

US20080213859A1 Methods for the Reduction of Stutter in Microsatellite Amplification 审中-公开
标题翻译：减少微卫星放大技术的方法
公开(公告)号：US20080213859A1
公开(公告)日：2008-09-04
申请号：US11742524
申请日：2007-04-30
申请人： Sulekha Rao Coticone , Will Bloch
发明人： Sulekha Rao Coticone , Will Bloch
IPC分类号： C12N9/10
CPC分类号： C12Q1/686 , C12Q1/6846 , C12Q1/6848 , C12Q1/6876 , C12Q2527/125 , C12Q2525/151
摘要： The invention provides a method for reducing stutter in the amplification of a microsatellite comprising the steps of providing a sample comprising a microsatellite having a G+C content of 50% or less; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with the enzyme for a sufficient amount of time and under conditions sufficient to amplify the microsatellite; wherein the incubation is performed in the presence of an amount of betaine, sorbitol or mixtures thereof, effective to reduce stutter relative to the amount of stutter observed in the absence of betaine and/or sorbitol. The invention also provides compositions containing betaine and/or sorbitol, kits for amplifying microsatellites having a G+C content of 50% or less, and methods of using all of the foregoing.
摘要翻译：本发明提供了减少微卫星扩增中的口吃的方法，包括以下步骤：提供包含G + C含量为50％或更少的微卫星的样品; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与酶温育足够的时间和足以扩增微卫星的条件; 其中在一定量的甜菜碱，山梨糖醇或其混合物的存在下进行培养，其有效地相对于在不存在甜菜碱和/或山梨醇的情况下观察到的口吃量减少口吃。本发明还提供含有甜菜碱和/或山梨糖醇的组合物，用于扩增G + C含量为50％以下的微卫星的试剂盒，以及使用上述所有方法。

17. 发明申请

US20070054287A1 METHOD FOR IDENTIFYING MEDICALLY IMPORTANT CELL POPULATIONS USING MICRO RNA AS TISSUE SPECIFIC BIOMARKERS 审中-公开
标题翻译：使用微RNA作为组织特异性生物标志物鉴定医学重要细胞群体的方法
公开(公告)号：US20070054287A1
公开(公告)日：2007-03-08
申请号：US11421456
申请日：2006-05-31
申请人： Will Bloch
发明人： Will Bloch
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/686 , C12Q1/6886 , C12Q2600/16 , C12Q2600/178 , C12Q2525/207
摘要： The present teachings provide methods for diagnosing biological conditions, including cancer. In some embodiments, a test sample is collected from a subject such as a clinical patient, wherein the test sample comprises background tissue and may or may not contain cells from a tissue of interest. Observation of a target miRNA normally present in a tissue of interest, but collected in an anatomical location ectopic to the tissue of interest, can be indicative of a biological condition. The present teachings further provide exponential amplification techniques applicable to performing these analyses.
摘要翻译：本教导提供了诊断包括癌症在内的生物学条件的方法。在一些实施方案中，从诸如临床患者的受试者收集测试样品，其中测试样品包含背景组织，并且可以包含或不包含来自感兴趣组织的细胞。通常存在于感兴趣的组织中但是收集在异位到感兴趣的组织的解剖位置的目标miRNA的观察可以指示生物学条件。本教导进一步提供适用于执行这些分析的指数扩增技术。

18. 发明授权

US06783940B2 Method of reducing non-specific amplification in PCR 有权
标题翻译：降低PCR中非特异性扩增的方法
公开(公告)号：US06783940B2
公开(公告)日：2004-08-31
申请号：US09998887
申请日：2001-10-31
申请人： Ian J. McLaughlin , Sulekha Rao Coticone , Will Bloch
发明人： Ian J. McLaughlin , Sulekha Rao Coticone , Will Bloch
IPC分类号： C12Q168
CPC分类号： C12Q1/686 , C12Q2527/125
摘要： The invention provides methods for reducing non-specific amplification DNA in a polymerase chain reaction comprising providing a sample comprising a target DNA sequence of interest; contacting the sample with at least one enzyme having nucleic acid polymerase activity; and incubating the sample with said enzyme for a time and under conditions sufficient to amplify the target DNA sequence, forming amplified target sequence; wherein the incubation is performed in the presence of an amount of sorbitol, or sorbitol and DMSO effective to reduce the non-specific amplification relative to the amount of non-specific amplification observed in the absence of sorbitol, or sorbitol and DMSO. The methods are suitable for amplification of ribosomal DNA, particularly from clinical samples. Compositions and kits containing sorbitol, or sorbitol and DMSO for reducing non-specific amplification are also provided.
摘要翻译：本发明提供了降低聚合酶链反应中的非特异性扩增DNA的方法，包括提供包含目标DNA序列的样品; 使样品与至少一种具有核酸聚合酶活性的酶接触; 并将样品与所述酶一起孵育一段时间并在足以扩增靶DNA序列的条件下形成扩增的靶序列; 其中在一定量的山梨糖醇或山梨糖醇和DMSO的存在下进行培养，所述山梨糖醇或山梨糖醇和DMSO相对于在不存在山梨醇或山梨糖醇和DMSO的情况下观察到的非特异性扩增量有效减少非特异性扩增。该方法适用于扩增核糖体DNA，特别是临床样品。还提供了含有山梨醇或山梨糖醇和DMSO用于降低非特异性扩增的组合物和试剂盒。

19. 发明授权

US5856192A Precision and accuracy of anion-exchange separation of nucleic acids 失效
标题翻译：核酸阴离子交换分离的精确度和精度
公开(公告)号：US5856192A
公开(公告)日：1999-01-05
申请号：US589904
申请日：1996-01-23
申请人： Will Bloch
发明人： Will Bloch
IPC分类号： B01D15/36 , B01J41/20 , G01N27/447 , G01N30/02 , G01N30/34 , G01N30/88
CPC分类号： B01D15/363 , B01J41/20 , G01N30/34 , G01N2030/8813 , G01N30/02 , Y10T436/108331 , Y10T436/143333
摘要： Solvents for salt-gradient anion-exchange separation of nucleic acids, especially double-stranded DNA and especially by liquid chromatography, are improved by replacing NaCl as the eluting salt with any of a wide range of alkyl ammonium salts and can be used to elute nucleic acids in strict order of increasing length, with reduced sensitivity to elution temperature and salt concentration. Anion-exchange chromatography with these solvents is well suited for identification of DNA fragments on the basis of size, with greater accuracy, precision, and resolvable size range than often is possible with gel electrophoresis.
摘要翻译：用于盐酸梯度阴离子交换分离核酸，特别是双链DNA，特别是通过液相色谱法的溶剂，通过用大范围的烷基铵盐代替NaCl作为洗脱盐，可以用于洗脱核酸酸按照长度增加的严格顺序，对洗脱温度和盐浓度的敏感性降低。使用这些溶剂的阴离子交换色谱法非常适合基于大小鉴定DNA片段，具有更高的准确度，精确度和可分辨的大小范围，通常可以通过凝胶电泳。

20. 发明授权

US5720923A Nucleic acid amplification reaction apparatus 失效
公开(公告)号：US5720923A
公开(公告)日：1998-02-24
申请号：US299033
申请日：1994-08-31
申请人： Lawrence A. Haff , Enrico Picozza , Will Bloch , Robert Ragusa , Joseph DiCesare , David Tracy , Paul Saviano , Timothy M. Woudenberg
发明人： Lawrence A. Haff , Enrico Picozza , Will Bloch , Robert Ragusa , Joseph DiCesare , David Tracy , Paul Saviano , Timothy M. Woudenberg
IPC分类号： C12N15/09 , B01J19/00 , B01L7/00 , C12M1/00 , C12Q1/68 , C40B40/06 , C40B60/14 , G01N35/00 , C12M1/40 , C12P19/34
CPC分类号： B01L7/525 , B01J19/0046 , B01L7/52 , B01L9/065 , B01J2219/00495 , B01J2219/00722 , B01L2300/1883 , C12Q1/686 , C40B40/06 , C40B60/14 , G01N2035/00237 , Y10T436/116664
摘要： Apparatus and method for performing a nucleic acid amplification reaction and preferably a polymerase chain reaction (PCR) in a reaction mixture in at least one capillary tube. Several different embodiments are disclosed. One embodiment cycles a sample through a capillary tube loop passing through two thermostatted fluid baths. Another embodiment has the capillary tube routed alternatingly between two heat exchangers to that the sample makes only one pass through the tube. Other embodiments maintain the heat exchangers stationary and translate the samples between them. Still further embodiments maintain the samples stationary and either automatically translate or rotate the heat exchangers past the samples contained within the capillary tubes to perform the thermal cycles necessary for the amplification reaction.

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