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    Methods and compositions for detection of small RNAs
    13.
    发明授权
    Methods and compositions for detection of small RNAs 有权

    公开(公告)号：US09416402B2

    公开(公告)日：2016-08-16

    申请号：US14600550

    申请日：2015-01-20

    申请人： SomaGenics, Inc.

    发明人： Sergei A. Kazakov ,  Pavan Kumar ,  Brian H. Johnston

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/682 ,  C12Q1/6806 ,  C12Q1/6844 ,  C12Q1/6865 ,  C12Q2525/313 ,  C12Q2521/501 ,  C12Q2525/143 ,  C12Q2525/207 ,  C12Q2531/125

    摘要： Currently, the circularization of small RNAs is broadly regarded as an obstacle in ligation-related assays and explicitly avoided while short lengths of linear RNA targets is broadly recognized as a factor limiting use of conventional primers in PCR-related assays. In contrast, the disclosed invention capitalizes on circularization of small RNA targets or their conjugates with oligonucleotide adapters. The circular RNA templates provide amplification of the target sequences via synthesis of multimer nucleic acids that can be either labeled for direct detection or subjected to PCR amplification and detection. Structure of small circular RNAs and corresponding multimeric nucleic acids provide certain advantages over current methods including flexibility in design of conventional RT and PCR primers as well as use of 5′-overlapping dimer-primers for efficient and sequence-specific amplification of short target sequences. Our invention also reduces number of steps and reagents while increasing sensitivity and accuracy of detection of small RNAs with both 2′OH and 2′-OMe at their 3′ ends. Our invention increase sensitivity and specificity of detection of microRNAs and other small RNAs with both 2′OH and 2′-OMe at their 3′ ends while allowing us to distinguish these two forms from each other.

    Methods and compositions for detection of small RNAs
    14.
    发明授权
    Methods and compositions for detection of small RNAs 有权
    用于检测小RNA的方法和组合物

    公开(公告)号：US08962253B2

    公开(公告)日：2015-02-24

    申请号：US13264122

    申请日：2010-04-13

    申请人： Sergei A. Kazakov ,  Pavan Kumar ,  Brian H. Johnston

    发明人： Sergei A. Kazakov ,  Pavan Kumar ,  Brian H. Johnston

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/682 ,  C12Q1/6806 ,  C12Q1/6844 ,  C12Q1/6865 ,  C12Q2525/313 ,  C12Q2521/501 ,  C12Q2525/143 ,  C12Q2525/207 ,  C12Q2531/125

    摘要： Currently, the circularization of small RNAs is broadly regarded as an obstacle in ligation-related assays and explicitly avoided while short lengths of linear RNA targets is broadly recognized as a factor limiting use of conventional primers in PCR-related assays. In contrast, the disclosed invention capitalizes on circularization of small RNA targets or their conjugates with oligonucleotide adapters. The circular RNA templates provide amplification of the target sequences via synthesis of multimer nucleic acids that can be either labeled for direct detection or subjected to PCR amplification and detection. Structure of small circular RNAs and corresponding multimeric nucleic acids provide certain advantages over current methods including flexibility in design of conventional RT and PCR primers as well as use of 5′-overlapping dimer-primers for efficient and sequence-specific amplification of short target sequences. Our invention also reduces number of steps and reagents while increasing sensitivity and accuracy of detection of small RNAs with both 2′OH and 2′-OMe at their 3′ ends. Our invention increase sensitivity and specificity of detection of microRNAs and other small RNAs with both 2′OH and 2′-OMe at their 3′ ends while allowing us to distinguish these two forms from each other.

    摘要翻译： 目前，小RNA的循环被广泛地认为是连接相关测定中的障碍，并被明确地避免，而短的线性RNA靶被广泛认为是限制PCR相关测定中常规引物的使用的因素。 相比之下，所公开的发明利用小RNA靶标或其与寡核苷酸衔接子的缀合物的环化。 循环RNA模板通过多聚核酸的合成提供靶序列的扩增，所述多聚核酸可以被标记用于直接检测或进行PCR扩增和检测。 小循环RNA和相应的多聚体核酸的结构提供了比现有方法（包括常规RT和PCR引物的设计灵活性）以及使用5'-重叠的二聚体 - 引物用于短目标序列的有效和序列特异性扩增的一些优点。 我们的发明还减少了步骤和试剂的数量，同时提高了在3'端检测2'OH和2'-OMe的小RNA的灵敏度和准确度。 我们的发明提高了在其3'末端检测微RNA和其他具有2'OH和2'-OMe的小RNA的灵敏度和特异性，同时允许我们将这两种形式相互区分开。

    Methods and compositions for detection of small RNAs
    17.
    发明授权
    Methods and compositions for detection of small RNAs 有权

    公开(公告)号：US10041107B2

    公开(公告)日：2018-08-07

    申请号：US15287254

    申请日：2016-10-06

    申请人： SomaGenics, Inc.

    发明人： Sergei A. Kazakov ,  Pavan Kumar ,  Brian H. Johnston

    IPC分类号： C12Q1/68 ,  C12Q1/682 ,  C12Q1/6806 ,  C12Q1/6865

    摘要： Currently, the circularization of small RNAs is broadly regarded as an obstacle in ligation-related assays and explicitly avoided while short lengths of linear RNA targets is broadly recognized as a factor limiting use of conventional primers in PCR-related assays. In contrast, the disclosed invention capitalizes on circularization of small RNA targets or their conjugates with oligonucleotide adapters. The circular RNA templates provide amplification of the target sequences via synthesis of multimer nucleic acids that can be either labeled for direct detection or subjected to PCR amplification and detection. Structure of small circular RNAs and corresponding multimeric nucleic acids provide certain advantages over current methods including flexibility in design of conventional RT and PCR primers as well as use of 5′-overlapping dimer-primers for efficient and sequence-specific amplification of short target sequences. Our invention also reduces number of steps and reagents while increasing sensitivity and accuracy of detection of small RNAs with both 2′OH and 2′-OMe at their 3′ ends. Our invention increase sensitivity and specificity of detection of microRNAs and other small RNAs with both 2′OH and 2′-OMe at their 3′ ends while allowing us to distinguish these two forms from each other.

    Methods and compositions for detection of small RNAS
    20.
    发明授权
    Methods and compositions for detection of small RNAS 有权
    用于检测小RNAS的方法和组合物

    公开(公告)号：US09493818B2

    公开(公告)日：2016-11-15

    申请号：US14835544

    申请日：2015-08-25

    申请人： SomaGenics, Inc.

    发明人： Sergei A. Kazakov ,  Pavan Kumar ,  Brian H. Johnston

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12Q1/682 ,  C12Q1/6806 ,  C12Q1/6844 ,  C12Q1/6865 ,  C12Q2525/313 ,  C12Q2521/501 ,  C12Q2525/143 ,  C12Q2525/207 ,  C12Q2531/125

    摘要： Currently, the circularization of small RNAs is broadly regarded as an obstacle in ligation-related assays and explicitly avoided while short lengths of linear RNA targets is broadly recognized as a factor limiting use of conventional primers in PCR-related assays. In contrast, the disclosed invention capitalizes on circularization of small RNA targets or their conjugates with oligonucleotide adapters. The circular RNA templates provide amplification of the target sequences via synthesis of multimer nucleic acids that can be either labeled for direct detection or subjected to PCR amplification and detection. Structure of small circular RNAs and corresponding multimeric nucleic acids provide certain advantages over current methods including flexibility in design of conventional RT and PCR primers as well as use of 5′-overlapping dimer-primers for efficient and sequence-specific amplification of short target sequences. Our invention also reduces number of steps and reagents while increasing sensitivity and accuracy of detection of small RNAs with both 2′OH and 2′-OMe at their 3′ ends. Our invention increase sensitivity and specificity of detection of microRNAs and other small RNAs with both 2′OH and 2′-OMe at their 3′ ends while allowing us to distinguish these two forms from each other.

    摘要翻译： 目前，小RNA的循环被广泛地认为是连接相关测定中的障碍，并被明确地避免，而短的线性RNA靶被广泛认为是限制PCR相关测定中常规引物的使用的因素。 相比之下，所公开的发明利用小RNA靶标或其与寡核苷酸衔接子的缀合物的环化。 循环RNA模板通过多聚核酸的合成提供靶序列的扩增，所述多聚核酸可以被标记用于直接检测或进行PCR扩增和检测。 小循环RNA和相应的多聚体核酸的结构提供了比现有方法（包括常规RT和PCR引物的设计灵活性）以及使用5'-重叠的二聚体 - 引物用于短目标序列的有效和序列特异性扩增的一些优点。 我们的发明还减少了步骤和试剂的数量，同时提高了在3'端检测2'OH和2'-OMe的小RNA的灵敏度和准确度。 我们的发明提高了在其3'末端检测微RNA和其他具有2'OH和2'-OMe的小RNA的灵敏度和特异性，同时允许我们将这两种形式相互区分开。