Method for quantitative detection of mercury ions through gold label silver stain and kit thereof
A technology of gold standard silver staining and quantitative detection, which is applied in the preparation of test samples, material analysis by observing the influence of chemical indicators, and analysis by making materials undergo chemical reactions, etc., to achieve high sensitivity, low price, Ease of pervasive effect
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Embodiment example 1
[0038] A method for the quantitative detection of mercury ions by gold-labeled silver staining of the present invention specifically comprises the following steps:
[0039] (1) Preparation of silanized glass slides
[0040] Place the glass slide in piranha washing solution, heat it at 90-100°C for 20-40 min, take it out, wash it three times with distilled water, and place it under N 2 After drying in the atmosphere, immerse in 1-3wt% 3-aminopropyltriethoxysilane (APTES) solution, silanize at room temperature for 2-6h, then wash with ethanol, vacuum at 110-130°C After drying for 1-3 hours, a silanized glass slide is obtained, and the piranha washing solution is obtained by mixing 98% concentrated sulfuric acid and 30% hydrogen peroxide at a volume ratio of 7:3;
[0041] (2) Preparation of specific DNA-nanogold probe solution
[0042] Mix 30-40 ul of 100 uM DNA with the sequence HS-5'-TTTTTTTTTT-3' and 0.5-1.5 mL of nano-gold solution with a concentration of 20-25 nM, shake we...
Embodiment example 2
[0055] The present invention is a kit for the quantitative detection of mercury ions by gold-labeled silver staining, comprising silanized glass slides, specific DNA-nano-gold probes and silver staining reagents, and the specific DNA-nano-gold probes are combined with nano-gold Specific DNA, the sequence of the specific DNA is as follows: HS-5'-TTTTTTTTTT-3'.
[0056] In this specific example, the diameter of the gold nanoparticle is 15-30 nm, and the structure of the specific DNA-nanogold probe is AuNPs-HS-5'-TTTTTTTTTT-3'. The silver staining reagent is composed of component A and component B, and the component A is AgNO containing 0.25 g / mL 3 Solution, component B is 0.0425 g / mL of hydroquinone, 0.064 g / mL of citric acid, and 0.059 g / mL of trisodium citrate mixed in an equal volume ratio. Before use, component A and component B Mix at a volume ratio of 3:100 to obtain the silver staining reagent.
[0057] In this specific example, the process of silanizing the glass slide...
Embodiment example 3
[0059] The establishment of the quantitative relationship between the gold-labeled silver-stained signal gray value and the concentration of mercury ions of the present invention specifically comprises the following steps:
[0060] (1) Silanization of glass slides
[0061] The slides were placed in piranha washing solution, heated at 90°C for 40 min, removed, washed three times with distilled water, and placed in N 2 After drying in the atmosphere, immerse in 2% 3-aminopropyltriethoxysilane (APTES) solution for silanization treatment at room temperature for 6 h, then wash with ethanol and then vacuum dry at 110 °C for 3 h. The above piranha washing solution is 98% concentrated sulfuric acid and 30% hydrogen peroxide are obtained by mixing in a volume ratio of 7: 3;
[0062] (2) Preparation of specific DNA-nanogold probe solution
[0063] Mix 34 μL of 100 μM DNA with the sequence HS-5'-TTTTTTTTTT-3' and 1 mL of 22.4 nM nano-gold solution, shake well, incubate at room temperat...
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