| 摘要 |
<p>PURPOSE: A method for detecting target nucleic acid sequences on a solid phase using a dual-labeled immobilized probe is provided to easily label a proper site of a probe with maximized quenching efficiency of a dual-labeling system, thereby minimizing background signals. CONSTITUTION: A method for detecting target nucleic acid sequences on a solid phase from a DNA or a nucleic acid mixture using a double-labeled probe with resistance to 5'->3' exonuclease activity of DNA polymerase comprises the steps of: hybridizing the target nucleic acid sequences on the immobilized probe; contacting the resultant with DNA polymerase under a condition when the immobilized probe is cleaved; finishing the cleavage of the exonuclease at a second label-labeled nucleotide; and detecting the change of a signal on the solid base. The change of the signal by the cleavage of the immobilized probe indicates the presence of the target nucleic acid sequences. [Reference numerals] (AA) Resistance to 5'-3' exonuclease activity of DNA polymerase; (BB) Target nucleic acid sequence; (CC) DNA polymerase with 5'-3' exonuclease activity; (DD) Double-labeled probe; (EE) Linker; (FF) Solid support; (GG) Nucleotide which is labeled has resistance to 5'-3' exonuclease activity of DNA polymerase, the label remains on the solid support, R is a fluorescent reporter molecule conjugated to the base of inner nucleotide, and Q is a quencher molecule conjugated to 5'-end of the double-labeled probe</p> |
