A kind of preparation method and application of silver-coated gold nanorods
A gold nanorod, nanorod technology, applied in the fields of nanostructure manufacturing, nanotechnology, nanotechnology, etc., can solve the problems of expensive instruments, cumbersome operation, high false positive and false negative rates of PCR, etc., and achieve the distribution of aspect ratio. Uniform, wide linear range, highly reproducible results
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Embodiment 1A
[0040] The synthesis of embodiment 1AuAg nanorods
[0041] First, weigh 0.364g CTAB, fully dissolve it in 20mL water, then add 10mL gold nanorod solution to the CTAB aqueous solution under stirring at 27°C, keep stirring for 2min, then add to the reaction system in the following order: 3mMAgNO 3 , 650 μL of 0.1M VC, 1200 μL of 0.1M NaOH, stirring continuously for 5-10 min and standing overnight, the reaction temperature is 27°C.
[0042] Note: Depending on the size of the synthesized AuAg nanorods, choose AgNO 3 The volume of the solution also varies, i.e. the choice of AgNO 3 The volumes of the solutions were 1250 μL, 2500 μL and 3750 μL, respectively.
[0043] figure 1 For Au nanorods and AuAg nanorods TEM (transmission electron microscope image). In the present invention, Au nanorods of different lengths ( figure 1 A, B, C) of silver-coated experiments were carried out, and AuAg nanorods of different lengths were obtained ( figure 1 D, E, F), according to the scale b...
Embodiment 2
[0044] Example 2 AuAg nanorods are used for eaeA gene detection
[0045] Schematic diagram of AuAg nanorod detection of Escherichia coli E.coliO157:H7eaeA gene figure 2 shown. The oligonucleotide probes were modified on the AuAg nanorods, and the fluorescent signal was enhanced after the oligonucleotide probes were hybridized with some nucleotides connected with the fluorescent dye cy3. When the target DNA appears, the probe is combined with the target DNA, and the fluorescence is weakened at this time, so it is applied to the detection of the E. coli O157:H7eaeA gene according to the principle of fluorescence enhancement-decrease.
[0046] The specific method is as follows:
[0047] (1) AuAg nanorod surface modification MUDA
[0048] First, the synthesized AuAg nanorods with a length of 50nm were centrifuged at 8500rpm at 25°C, and then, according to the principle of ligand exchange reaction, the surface was modified with mercaptoundecanoic acid (MUDA), and 3 mL of purifi...
Embodiment 3
[0060] Performance analysis of embodiment 3AuAg nanorods
[0061] (1) Stability
[0062] Different batches of AuAg nanorods prepared by the present invention were used to detect the eaeA gene of Escherichia coli E. coliO157:H7. The results are shown in Table 1. The result shows that the gene recovery rate is 98.36-101.67%, and the error is small, indicating that the AuAg nanorod and the detection method of the present invention have good stability and repeatability.
[0063] Table 1 Detection of Escherichia coli E.coliO157:H7eaeA gene recovery rate and error results
[0064]
[0065] Note: This experiment is a recovery experiment, the background concentration is consistent, and the variable is the spiked concentration.
[0066] (2) specificity
[0067] Using the AuAg nanorods prepared in the present invention, detection contains different gene sequences, including the target sequence (5'-CGTCGCGGTATAAGTAATGGTATCGGCGTT-3', which is completely complementary to the oligonu...
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