DNA hydrogel based on biological mimic enzyme signal amplification and application thereof
A biosimulation and hydrogel technology, applied in the direction of chemical reaction analysis, chemical/physical process, physical/chemical process catalyst, etc., can solve food safety hazards and other problems, and achieve low cost and good portability , detect the effect of fast
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Embodiment 1
[0060] This example prepares a DNA hydrogel based on biomimetic enzyme signal amplification, in which the biomimetic enzyme is Cu / Au / Pt with peroxidase activity, and the outer hydrogel coating uses linear polymer The backbone of the material is a straight-chain polyacrylamide backbone. The following is the preparation method of the DNA hydrogel of this embodiment:
[0061] Step 1: Preparation method of Cu / Au / Pt TNPs:
[0062] Cu / Au / Pt TNPs biomimetic enzymes with peroxidase activity were synthesized by a "one-pot method". 12 μL CuSO 4 ﹒ 5H 2 O (0.1mol / L) and 25μL trisodium citrate (0.1mol / L) were added to 10mL ultrapure water, mixed well and then added 500μL KBH 4 (25mmol / L), after stirring for 15min, 25μL HAuCl 4 (0.1mol / L) and 25μL K 2 PtCl 4 (0.1mol / L) were added dropwise to the above mixture, and Cu / Au / Pt TNPs could be obtained after stirring for 20 min.
[0063] Step 2: Design of the first short-strand DNA and the second short-strand DNA
[0064] Design principl...
Embodiment 2
[0076] In this example, whether the binding competitiveness between the first short-strand DNA (SA) and the second short-strand DNA (SB) and the aptamer designed in Example 1 satisfies the points ①-② in the design principle verify. The verification method is as follows:
[0077] (1) Sample preparation:
[0078]
[0079] Note: Before adding MC-LR, denature the mixture at 95°C for 5 minutes, cool it to room temperature and place it at 4°C for 20 minutes. After adding MC-LR, incubate at 37°C for 40 minutes and store it in a -20°C refrigerator for testing.
[0080] (2) Take 200 μL of the samples in the above table respectively in a cuvette, and put them on a circular dichroism spectrometer for measurement. Circular dichroism instrument parameters are set as follows: measurement wavelength range: 200-350nm, scanning speed 100nm / min, response time 4s, bandwidth 2.0nm, scanning mode: continuous. Baseline correction: PBS solution.
[0081] (3) Data analysis:
[0082] Import th...
Embodiment 3
[0089] In this example, the DNA hydrogel based on the signal amplification of the biomimetic enzyme prepared in Example 1 is used for the quantitative detection of MC-LR in water, and the detection method is:
[0090] Take 10 μL of the Cu / Au / Pt-DNA hydrogel prepared in Example 1 into a 1.5 mL disposable centrifuge tube, add the sample solution to be tested (including MC-LR), and incubate at 37° C. for 40 min. Add 198 μL HO to the mixed system 2 O (pH=3), and the final concentrations of TMB and 48mmol / L of H 2 o 2 , After reacting at room temperature, concentrated hydrochloric acid was added to terminate the reaction. The absorbance value at a specific wavelength (optimally 452 nm) is measured by a UV-visible spectrophotometer.
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