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    • 6. 发明申请
    • METHODS FOR ENHANCING NUCLEIC ACID HYBRIDIZATION
    • 增加核酸杂交的方法
    • WO2011120049A1
    • 2011-09-29
    • PCT/US2011/030215
    • 2011-03-28
    • INTEGRATED DNA TECHNOLOGIES, INC.ROSE, ScottBEHLKE, Mark, A.OWCZARZY, RichardWALDER, Joseph, A.THOMAS, Derek, M.MARVIN, Michael, R.
    • ROSE, ScottBEHLKE, Mark, A.OWCZARZY, RichardWALDER, Joseph, A.THOMAS, Derek, M.MARVIN, Michael, R.
    • C12Q1/68A61K31/11
    • C12Q1/6832C07H21/04C12Q1/6818C12Q1/6851G01N33/5308G01N33/542C12Q2525/185C12Q2525/197C12Q2565/101
    • A composition, comprising a first oligonucleotide having the structure 5'-Y 1- L 1- X-L 2 _2-Y 2 -3' Y 1 comprises a sequence of four or more DNA or RNA nucleotides, including a first nucleotide N 1 having a 3' phosphate covalently linked to L 1. Y 2 comprises a sequence of four or more DNA or RNA nucleotides, including a second nucleotide N 2 having a 5' phosphate covalently linked to L 2. L 1 and L 2 each independently are a direct bond or a C 1 C 7 alkyl, alkynyl, alkenyl, heteroalkyl, substituted alkyl, aryl, heteroaryl, substituted aryl, cycloalkyl, alkylaryl, or alkoxyl group. X is R\ is a hydrogen or a d-Cg alkyl. M is a quencher. The first oligonucleotide is adapted to hybridize to a second oligonucleotide having the structure 3'-Υ3-Υ4-5'. Y3 comprises a sequence of four or more DNA or RNA nucleotides, including a third nucleotide N3. Y4 comprises a sequence of four or more DNA or RNA nucleotides, including a fourth nucleotide N4 that is directly attached to nucleotide N 3 . If the first oligonucleotide hybridizes to the second oligonucleotide, Ni base pairs with N 3 and N2 base pairs with N4.to form a duplex having a T m that is greater than the T m of a duplex formed between the second oligonucleotide and a third oligonucleotide having the structure 5'-Y 1 -Y 2 - 3'.
    • 包含结构为5'-Y1-L1-X-L2-2-Y2-3'Y1的第一寡核苷酸的组合物包含四个或更多个DNA或RNA核苷酸的序列,包括具有3'磷酸共价连接的第一个核苷酸N1 到L1。 Y2包含四个或更多个DNA或RNA核苷酸的序列,包括具有与L2共价连接的5'磷酸二酯的第二核苷酸N2。 L 1和L 2各自独立地是直接键或C 1 -C 7烷基,炔基,烯基,杂烷基,取代的烷基,芳基,杂芳基,取代的芳基,环烷基,烷基芳基或烷氧基。 X是R 1是氢或C 1 -C 8烷基。 M是猝灭剂。 第一寡核苷酸适于与具有结构3'-β3-3→4-5'的第二寡核苷酸杂交。 Y3包含四个或更多个DNA或RNA核苷酸的序列,包括第三核苷酸N3。 Y4包含四个或更多个DNA或RNA核苷酸的序列,其包括与核苷酸N3直接相连的第四核苷酸N4。 如果第一寡核苷酸与第二寡核苷酸杂交,则具有N 3和N 2碱基对的Nε碱基对与N4.形成具有比第二寡核苷酸和具有结构的第三寡核苷酸之间形成的双链体的Tm更大的Tm的双链体 5'-Y1-Y2-3' 。