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1. 发明申请

US20060177867A1 Repair of nucleic acids for improved amplification 有权
标题翻译：修复核酸以改善扩增
公开(公告)号：US20060177867A1
公开(公告)日：2006-08-10
申请号：US11401826
申请日：2006-04-11
申请人： Thomas Evans , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan , Rebecca Kucera
发明人： Thomas Evans , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan , Rebecca Kucera
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6844 , C12P19/34 , C12Q1/6848 , C12Q1/686 , C12Q2521/101 , C12Q2521/501 , C12Q2521/331 , C12Q2521/301 , C12Q2521/514
摘要： Methods and compositions are provided for repairing a polynucleotide so that it can be copied with improved fidelity and/or yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NAD+ or ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI. The reaction mixture may further contain an AP endonuclease and a polymerase. If used, these enzymes may be selected according to their ability to withstand high temperatures. For example, the reaction mixture may be used prior to a polynucleotide synthesis reaction in which case enzymes that are not thermophilic may be used. The repair reaction is not time sensitive with respect to seconds, minutes or hours of incubation in the enzyme mixture in as much as the repair is effected rapidly and prolonged incubation is not generally adverse.
摘要翻译：提供了用于修复多核苷酸的方法和组合物，使得其可以在例如扩增反应中以改进的保真度和/或产率复制。这包括使用包含连接酶和选自NAD +或ATP的辅因子的反应混合物，并在不存在核酸内切酶VI的情况下将多核苷酸与反应混合物一起温育。反应混合物还可以含有AP内切核酸酶和聚合酶。如果使用，这些酶可以根据其耐受高温的能力来选择。例如，反应混合物可以在多核苷酸合成反应之前使用，在这种情况下可以使用不是嗜热的酶。维修反应对于酶混合物中的几秒钟，几分钟或几小时的孵育时间并不敏感，只要修复迅速进行并且延长的孵育通常不是不利的。

2. 发明申请

US20100173364A1 Repair of Nucleic Acids for Improved Amplification 审中-公开
标题翻译：用于改进扩增的核酸修复
公开(公告)号：US20100173364A1
公开(公告)日：2010-07-08
申请号：US12296827
申请日：2007-04-11
申请人： Thomas C. Evans, JR. , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan , Rebecca Kucera
发明人： Thomas C. Evans, JR. , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan , Rebecca Kucera
IPC分类号： C12P19/34 , C12N9/00 , C12N9/10
CPC分类号： C12Q1/686 , C12Q2521/514 , C12Q2521/501 , C12Q2521/301 , C12Q2525/119
摘要： Methods and compositions are provided for repairing a polynucleotide so that it can be copied with improved fidelity and/o yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a DNA ligase and an effective amount of at least one endonuclease as well as a cofactor selected from NAD+ or ATP.
摘要翻译：提供了用于修复多核苷酸的方法和组合物，使得其可以在例如扩增反应中以改进的保真度和/或产率复制。这包括使用包括DNA连接酶和有效量的至少一种内切核酸酶以及选自NAD +或ATP的辅因子的反应混合物。

3. 发明申请

US20060088868A1 Repair of nucleic acids for improved amplification 有权
标题翻译：修复核酸以改善扩增
公开(公告)号：US20060088868A1
公开(公告)日：2006-04-27
申请号：US11255290
申请日：2005-10-20
申请人： Thomas Evans , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan
发明人： Thomas Evans , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/686 , C12Q1/6844 , C12Q1/6848 , C12Q2521/501 , C12Q2521/301 , C12Q2521/514 , C12Q2521/101 , C12Q2521/331
摘要： Methods and compositions are provided for repairing a polynucleotide so that it can be synthesized efficiently with improved fidelity and yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NAD+ or ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI. The reaction mixture may further contain an AP endonuclease and a polymerase. These enzymes are optionally selected according to their ability to withstand high temperatures so they can be included in an amplification mixture. The reaction mixture may be used prior to a polynucleotide synthesis reaction in which case enzymes that are not thermophilic may be used. The repair reaction is not time sensitive with respect to seconds, minutes or hours of incubation in the enzyme mixture.
摘要翻译：提供了用于修复多核苷酸的方法和组合物，使得其可以在例如扩增反应中以提高的保真度和产率被有效地合成。这包括使用包含连接酶和选自NAD +或ATP的辅因子并在不存在核酸内切酶VI的情况下将多核苷酸与反应混合物一起温育的反应混合物。反应混合物还可以含有AP内切核酸酶和聚合酶。这些酶根据其耐受高温的能力任选地进行选择，因此它们可以被包括在扩增混合物中。反应混合物可以在多核苷酸合成反应之前使用，在这种情况下可以使用不是嗜热的酶。相对于在酶混合物中孵育的秒，分钟或数小时，修复反应不是时间敏感的。

4. 发明授权

US07700283B2 Repair of nucleic acids for improved amplification 有权
标题翻译：修复核酸以改善扩增
公开(公告)号：US07700283B2
公开(公告)日：2010-04-20
申请号：US11255290
申请日：2005-10-20
申请人： Thomas C. Evans , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan
发明人： Thomas C. Evans , Barton Slatko , Lixin Chen , Romaldas Vaisvila , Chudi Guan
IPC分类号： C12Q1/68
CPC分类号： C12Q1/686 , C12Q1/6844 , C12Q1/6848 , C12Q2521/501 , C12Q2521/301 , C12Q2521/514 , C12Q2521/101 , C12Q2521/331
摘要： Methods and compositions are provided for repairing a polynucleotide so that it can be synthesized efficiently with improved fidelity and yield in, for example, an amplification reaction. This involves the use of a reaction mixture that includes a ligase and a cofactor selected from NAD+ or ATP and incubating the polynucleotide with the reaction mixture in the absence of Endonuclease VI.The reaction mixture may further contain an AP endonuclease and a polymerase. These enzymes are optionally selected according to their ability to withstand high temperatures so they can be included in an amplification mixture. The reaction mixture may be used prior to a polynucleotide synthesis reaction in which case enzymes that are not thermophilic may be used. The repair reaction is not time sensitive with respect to seconds, minutes or hours of incubation in the enzyme mixture.
摘要翻译：提供了用于修复多核苷酸的方法和组合物，使得其可以在例如扩增反应中以提高的保真度和产率被有效地合成。这包括使用包含连接酶和选自NAD +或ATP的辅因子并在不存在核酸内切酶VI的情况下将多核苷酸与反应混合物一起温育的反应混合物。反应混合物还可以含有AP内切核酸酶和聚合酶。这些酶根据其耐受高温的能力任选地进行选择，因此它们可以被包括在扩增混合物中。反应混合物可以在多核苷酸合成反应之前使用，在这种情况下可以使用不是嗜热的酶。相对于在酶混合物中孵育的秒，分钟或数小时，修复反应不是时间敏感的。

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