Fluorescence precise quantification method of integrity of DNA origami structure
A technology of origami structure and integrity, applied in the field of nano-biological materials and biological detection, can solve the problems of being easily affected by the environment, structural stability, structural dissociation, etc., and achieve the effect of accurate quantitative method and easy evaluation operation
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Embodiment 1
[0025] The DNA triangular origami structure synthesized above was purified in an ultrafiltration centrifuge tube at a speed of 3000 g for 10 minutes, and centrifuged 3 times to remove excess staple chains.
[0026] (1) Set the final concentration of the purified DNA triangula to 2nM, take 10 μl of the DNA triangula solution, add 5 μl of 20×Eva Green, and add 1×TAE-Mg 2+ 85 μl of buffer solution, measure its fluorescence intensity, the structure remains intact (0%);
[0027] (2) Control Mg 2+ Concentration is 1 / 10 in the original buffer solution, stand for 5 minutes, measure its fluorescence intensity, the structure changes by 28.36%;
[0028] (3) Restore the Mg in the solution 2+ Concentration is 1×TAE-Mg 2+ , the fluorescence intensity was measured again, and the structure recovered to 0.88%, with figure 2 a.
[0029] Always keep the concentration of DNA triangular origami at 0.2 nM. Determine the completeness of the DNA origami structure according to the size of the f...
Embodiment 2
[0031] The DNA triangular origami structure synthesized above was purified in an ultrafiltration centrifuge tube at a speed of 3000 g for 10 minutes, and centrifuged 3 times to remove excess staple chains.
[0032] (1) Set the final concentration of the purified DNA triangula to 2nM, take 10 μl of the DNA triangula solution, add 5 μl of 20×Eva Green, and add 1×TAE-Mg 2+ 85 μl of buffer solution, measure its fluorescence intensity;
[0033] (2) Control Mg 2+ The concentration is 1 / 10 of the original buffer solution, let it stand for 1 hour, and measure its fluorescence intensity, the structure has changed by 33.45%;
[0034] (3) Restore the Mg in the solution 2+ Concentration is 1×TAE-Mg 2+ , measured its fluorescence intensity again, the structure could not be recovered, and the disturbance of the concentration of the recovery solution caused a greater damage to the structure to 37.91%. figure 2 b.
[0035] Always keep the concentration of DNA triangular origami at 0.2 n...
Embodiment 3
[0037] The DNA triangular origami structure synthesized above was purified in an ultrafiltration centrifuge tube at a speed of 3000 g for 10 minutes, and centrifuged 3 times to remove excess staple chains.
[0038] (1) Set the final concentration of the purified DNA triangula to 2nM, take 10 μl of the DNA triangula solution, add 5 μl of 20×Eva Green, and add 1×TAE-Mg 2+ 85 μl of buffer solution, measure its fluorescence intensity;
[0039] (2) Control Mg 2+ The concentration is 1 / 10 of the original buffer solution, let it stand for 5 minutes, measure its fluorescence intensity, and the structure changes by 33.03%;
[0040] (3) Add a small amount of Q water (10μl), observe the effect of solution disturbance on the fluorescence intensity, measure the fluorescence intensity again, the structure changes by 32.17%, attached figure 2 left picture.
[0041] Always keep the concentration of DNA triangular origami at 0.2 nM. Determine the completeness of the DNA origami structure ...
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