公开(公告)号：US06977148B2

公开(公告)日：2005-12-20

申请号：US09977868

申请日：2001-10-15

申请人： Frank B. Dean ,  Roger S. Lasken ,  Linhua Fang ,  A. Fawad Faruqi ,  Osama A. Alsmadi ,  Mark D. Driscoll ,  Seiyu Hosono

发明人： Frank B. Dean ,  Roger S. Lasken ,  Linhua Fang ,  A. Fawad Faruqi ,  Osama A. Alsmadi ,  Mark D. Driscoll ,  Seiyu Hosono

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/6844 ,  C12Q2527/125

摘要： Disclosed are compositions and a method for amplification of nucleic acid sequences of interest. The disclosed method generally involves replication of a target sequence such that, during replication, the replicated strands are displaced from the target sequence by strand displacement replication of another replicated strand. In one form of the disclosed method, the target sample is not subjected to denaturing conditions. It was discovered that the target nucleic acids, genomic DNA, for example, need not be denatured for efficient multiple displacement amplification. The primers used can be hexamer primers. The primers can also each contain at least one modified nucleotide such that the primers are nuclease resistant. The primers can also each contain at least one modified nucleotide such that the melting temperature of the primer is altered relative to a primer of the same sequence without the modified nucleotide(s). The DNA polymerase can be φ29 DNA polymerase.

摘要翻译： 公开了用于扩增感兴趣的核酸序列的组合物和方法。 所公开的方法通常涉及靶序列的复制，使得在复制期间，复制的链通过另一复制链的链置换复制而从靶序列位移。 在所公开的方法的一种形式中，目标样品不经历变性条件。 已经发现，靶核酸，例如基因组DNA不需要被变性以用于有效的多位置扩增。 所使用的引物可以是六聚体引物。 引物还可以各自含有至少一个修饰的核苷酸，使得引物是耐核酸酶的。 引物还可以各自包含至少一个修饰的核苷酸，使得引物的融解温度相对于没有修饰的核苷酸的相同序列的引物而改变。 DNA聚合酶可以是phi29 DNA聚合酶。

公开(公告)号：US09487823B2

公开(公告)日：2016-11-08

申请号：US10327602

申请日：2002-12-20

申请人： Roger S. Lasken ,  Michael Egholm ,  Osama A. Alsmadi

发明人： Roger S. Lasken ,  Michael Egholm ,  Osama A. Alsmadi

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6844 ,  C12Q1/6858 ,  C12Q2531/119 ,  C12Q2525/151

摘要： Disclosed are compositions and a method for amplification of nucleic acid sequences of interest. The disclosed method generally involves replication of a complex nucleic acid sample such as genomic samples using one, a few, or more primers such that, during replication, the replicated strands are displaced from the nucleic acid molecules in the sample by strand displacement replication of another replicated strand. It was discovered that highly complex nucleic acid samples can be efficiently amplified using only one or a few primers having specific nucleic acid sequences. The one or few primers are complementary to nucleic acid sequences distributed throughout nucleic acid in the sample.

摘要翻译： 公开了用于扩增感兴趣的核酸序列的组合物和方法。 所公开的方法通常涉及使用一种，几种或多种引物复制复杂核酸样品，例如基因组样品，使得在复制期间，复制的链通过另一个的链置换复制从样品中的核酸分子中移位 复制链。 已经发现，仅使用具有特定核酸序列的一个或几个引物可以有效扩增高度复杂的核酸样品。 一个或几个引物与分析在样品中的核酸的核酸序列互补。

公开(公告)号：US07074600B2

公开(公告)日：2006-07-11

申请号：US10272465

申请日：2002-10-15

申请人： Frank B. Dean ,  Roger S. Lasken ,  Linhua Fang ,  A. Fawad Faruqi ,  Osama A. Alsmadi ,  Mark D. Driscoll ,  Seiyu Hosono ,  Michele Wisniewski ,  Wanmin Song

发明人： Frank B. Dean ,  Roger S. Lasken ,  Linhua Fang ,  A. Fawad Faruqi ,  Osama A. Alsmadi ,  Mark D. Driscoll ,  Seiyu Hosono ,  Michele Wisniewski ,  Wanmin Song

IPC分类号： C12P19/34

CPC分类号： C12Q1/6844 ,  C12Q1/6806 ,  C12Q2527/125 ,  C12Q2527/119 ,  C12Q2537/143 ,  C12Q2531/119 ,  C12Q2525/125

摘要： Disclosed are compositions and a method for amplification of nucleic acid sequences of interest. The disclosed method generally involves replication of a target sequence such that, during replication, the replicated strands are displaced from the target sequence by strand displacement replication of another replicated strand. In one form of the disclosed method, the target sample is not subjected to denaturing conditions. It was discovered that the target nucleic acids, genomic DNA, for example, need not be denatured for efficient multiple displacement amplification. The primers used can be hexamer primers. The primers can also each contain at least one modified nucleotide such that the primers are nuclease resistant. The primers can also each contain at least one modified nucleotide such that the melting temperature of the primer is altered relative to a primer of the same sequence without the modified nucleotide(s). The DNA polymerase can be φ29 DNA polymerase.

摘要翻译： 公开了用于扩增感兴趣的核酸序列的组合物和方法。 所公开的方法通常涉及靶序列的复制，使得在复制期间，复制的链通过另一复制链的链置换复制而从靶序列位移。 在所公开的方法的一种形式中，目标样品不经历变性条件。 已经发现，靶核酸，例如基因组DNA不需要被变性以用于有效的多位置扩增。 所使用的引物可以是六聚体引物。 引物还可以各自含有至少一个修饰的核苷酸，使得引物是耐核酸酶的。 引物还可以各自包含至少一个修饰的核苷酸，使得引物的融解温度相对于没有修饰的核苷酸的相同序列的引物而改变。 DNA聚合酶可以是phi29 DNA聚合酶。

公开(公告)号：US07129087B2

公开(公告)日：2006-10-31

申请号：US10398833

申请日：2001-10-11

申请人： Fred R. Kramer ,  Osama A. Alsmadi ,  Sanjay Tyagi

发明人： Fred R. Kramer ,  Osama A. Alsmadi ,  Sanjay Tyagi

IPC分类号： C12N15/02 ,  C12N15/88 ,  A61K9/127

CPC分类号： A61K9/127 ,  A61K9/1271

摘要： Coalescence of cells or other membrane-bound entities is facilitated by anchoring an outwardly projecting first oligonucleotide in one member and an outwardly projecting second oligonucleotide, complementary to the first, in a second member and incubating under hybridizing conditions. Liposomes may be coalesced with cells to deliver hydrophilic agents thereto, such as DNA probes or drugs. Kits containing complementary oligonucleotides containing hydrophobic anchoring moieties may be used.

摘要翻译： 通过将向外突出的第一寡核苷酸锚定在一个构件中并通过在第二构件中与第一寡核苷酸互补的向外突出的第二寡核苷酸并在杂交条件下孵育来促进细胞或其它膜结合实体的聚结。 脂质体可以与细胞聚结以向其递送亲水剂，例如DNA探针或药物。 可以使用含有含有疏水性锚定部分的互补寡核苷酸的试剂盒。

公开(公告)号：US06573051B2

公开(公告)日：2003-06-03

申请号：US09803713

申请日：2001-03-09

申请人： Osama A. Alsmadi ,  Patricio Abarzua

发明人： Osama A. Alsmadi ,  Patricio Abarzua

IPC分类号： C12Q168

CPC分类号： C12Q1/6844 ,  C12Q2525/301 ,  C12Q2525/307 ,  C12Q2531/125

摘要： Disclosed are compositions and methods for reducing or eliminating generation of unwanted, undesirable, or non-specific amplification products in nucleic acid amplification reactions, such as rolling circle amplification. One form of composition is an open circle probe that can form an intramolecular stem structure, such as a hairpin structure, at one or both ends. The stem structure allows the open circle probe to be circularized when hybridized to a legitimate target sequence but results in inactivation of uncircularized open circle probes. This inactivation, which preferably involves stabilization of the stem structure, extension of the end of the open circle probe, or both, reduces or eliminates the ability of the open circle probe to prime nucleic acid synthesis or to serve as a template for rolling circle amplification. The disclosed method is useful for detection, quantitation, and/or location of any desired analyte, such as proteins and peptides.

摘要翻译： 公开了用于减少或消除在核酸扩增反应中产生不想要的，不期望的或非特异性扩增产物的组合物和方法，例如滚环扩增。 组合物的一种形式是可以在一端或两端形成分子内结构，如发夹结构的开环探针。 茎结构允许当与合法靶序列杂交时，开环探针被环化，但导致未圆形的开放圆探针失活。 这种失活优选涉及茎结构的稳定化，开环探针的末端的延伸或两者都减少或消除了开环探针引发核酸合成或用作滚环扩增的模板的能力 。 所公开的方法可用于任何所需分析物如蛋白质和肽的检测，定量和/或定位。