Real-time fluorescence quantitative PCR detection method and kit based on metal ruthenium complex
AssigneeShaanxi University of Science and Technology
InventorsQinfeng Xu, Jing Dong, Hongxin Song, Jianlan Liu
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Abstract
The present disclosure discloses a real-time fluorescence quantitative polymerase chain reaction (PCR) detection method and kit based on a metal ruthenium complex. The present disclosure is capable of establishing the detection method for performing a real-time fluorescence quantitative PCR by using the metal ruthenium complex as fluorescence dye.
Claims8
A real-time fluorescence quantitative PCR detection method based on a metal ruthenium complex, wherein the real-time fluorescence quantitative PCR detection method comprises the following steps:
- using a metal ruthenium complex as a dye component in a real-time fluorescence quantitative PCR reaction system;
- using multiple nucleic acid samples with different copy numbers as standard substances, using the each standard substance as an amplification template component in the real-time fluorescence quantitative PCR reaction system respectively, and performing a real-time fluorescence quantitative PCR according to a preset reaction program, after reaction is finished respectively, drawing a standard curve according to a cycle threshold of the each standard substance;
- using a nucleic acid sample with an unknown copy number as an amplification template component in the real-time fluorescence quantitative PCR reaction system, performing the real-time fluorescence quantitative PCR according to the reaction program, and calculating a DNA copy number of the sample according to the standard curve and a cycle threshold of the nucleic acid sample with the unknown copy number;
- wherein the metal ruthenium complex is [Ru(phen)2(dppz)]2+ or [Ru(bpy)2(dppz)]2+;
- wherein a final volume of the reaction system is 10-20 μL, and a final concentration of the metal ruthenium complex in the reaction system is 3-6 μM.
A real-time fluorescence quantitative PCR detection kit based on a metal ruthenium complex, wherein the kit comprises a metal ruthenium complex served as nucleic acid dye in a real-time fluorescene quantitative PCR reaction system,
- wherein the metal ruthenium complex is [Ru(phen)2(dppz)]2+ or [Ru(bpy)2(dppz)]2+;
- wherein a final volume of the reaction system is 10-20 μL, and a final concentration of the metal ruthenium complex in the reaction system is 3-6 μM.
A real-time fluorescence quantitative PCR detection system based on a metal ruthenium complex, wherein comprising a kit and a qPCR meter for preparing a real-time fluorescence quantitative PCR reaction system, the kit comprises a metal ruthenium complex served as nucleic acid dye in a real-time fluorescence quantitative PCR reaction system, while the qPCR machine meter performs fluorescence signal collection in an extending step of a circular reaction, a used collection channel of a fluorescence signal is as follows:
- an excitation wavelength is 400-500 nm, and an emission wavelength is 550-750 nm,
- wherein the metal ruthenium complex is [Ru(phen)2(dppz)]2+ or [Ru(bpy)2(dppz)]2+;
- wherein a final volume of the reaction system is 10-20 μL, and a final concentration of the metal ruthenium complex in the reaction system is 3-6 μM.
Dependencies are inferred from the claim text.