Method for screening G-quadruplex ligand based on DNA silver nano-cluster homogeneous phase
A technology of silver nanoclusters and quadruplexes, which is applied in the field of homogeneous screening of G-quadruplex ligands based on DNA-silver nanoclusters, can solve the problems of complicated operation, high cost, harsh experimental conditions, etc., and achieves universality. Strong, responsive, and easy-to-use effects
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Embodiment 1
[0022] Taking aloe-emodin derivative 3 (AED3) as an example, the method for screening whether it is a G-quadruplex ligand consists of the following steps:
[0023] (1) Prepare a tris-acetic acid buffer solution with a concentration of 10mmol / L pH=8.0 according to a conventional method, and use this buffer solution to separate the single-stranded nucleic acid probe (12C5TG, the sequence is: 5'-CCCCCCCCCCCCTTTTTTTAGGGTTAGGGTTAGGGTTAGGG-3 ') to prepare a stock solution with a concentration of 100 μmol / L for later use; add 40 μL 100 μmol / L single-stranded nucleic acid probe stock solution to 155.2 μL 10 mmol / L pH=8.0 tris-acetic acid buffer solution, Add 2.4μL 10mmol / L silver nitrate solution to the diluted nucleic acid probe solution, incubate on ice for 15 minutes, then quickly add 2.4μL 10mmol / L sodium borohydride to the above mixed solution to make the single-stranded nucleic acid probe in the final mixture The concentration of single-stranded nucleic acid probe, silver nitrat...
Embodiment 2
[0030] Taking aloe-emodin derivative 3 (AED3) as an example, the method for screening whether it is a G-quadruplex ligand consists of the following steps:
[0031] (1) Prepare a tris-acetic acid buffer solution with a concentration of 10mmol / L pH=8.0 according to a conventional method, and use this buffer solution to separate the single-stranded nucleic acid probe (12C5TG, the sequence is: 5'-CCCCCCCCCCCCTTTTTTTAGGGTTAGGGTTAGGGTTAGGG-3 ') to prepare a stock solution with a concentration of 100 μmol / L for later use; then add 40 μL 100 μmol / L single-stranded nucleic acid probe stock solution to 150.4 μL 10 mmol / L pH=8.0 tris-acetic acid buffer solution, and Dilute the single-stranded nucleic acid probe stock solution, add 4.8μL 10mmol / L silver nitrate solution to the diluted solution, incubate on ice for 10 minutes, quickly add 4.8μL 10mmol / L sodium borohydride to the above mixed solution to make the single-stranded nucleic acid probe The concentration of the needle in the final...
Embodiment 3
[0035] Taking aloe-emodin derivative 3 (AED3) as an example, the method for screening whether it is a G-quadruplex ligand consists of the following steps:
[0036] (1) Prepare a tris-acetic acid buffer solution with a concentration of 10mmol / L pH=8.0 according to a conventional method, and use this buffer solution to separate the single-stranded nucleic acid probe (12C5TG, the sequence is: 5'-CCCCCCCCCCCCTTTTTTTAGGGTTAGGGTTAGGGTTAGGG-3 ') to prepare a stock solution with a concentration of 100 μmol / L for later use; then add 40 μL 100 μmol / L single-stranded nucleic acid probe stock solution to 156 μL 10 mmol / L pH=8.0 tris-acetic acid buffer solution, and Dilute the strand nucleic acid probe stock solution, add 2.0 μL 10 mmol / L silver nitrate solution to the diluted solution, incubate on ice for 10 minutes, quickly add 2.0 μL 10 mmol / L sodium borohydride to the above mixed solution to make the single-stranded nucleic acid probe The concentration in the final mixture reaches 20 μ...
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