Fluorescent quantum dot/nano-metal particle conjugate and preparation and application thereof
A technology of nano metal particles and fluorescent quantum dots, applied in the field of nanomaterials, can solve problems such as expensive and lengthy sample preparation
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Embodiment 1
[0062] Example 1: Preparation of single-stranded DNA probe-modified gold nanoparticles
[0063] The synthesis of gold nanoparticles can be found in the methods of the prior art (J.J.Storhoff, R.Elghanian, R.C.Mucic, C.A.Mirkin, R.L.Letsinger.J.Am.Chem.Soc., 1998, 120, 1959-1964). Add 98 ml of water to a two-necked bottle. Add 2 ml of 50 mM HAuCl 4 The solution makes HAuCl in the bottle 4 The concentration was 1 mM, heated to reflux with stirring, then 10 ml of 38.8 mM sodium citrate solution was rapidly added, and the color changed from light yellow to dark red within 1 min. Reflux was continued for 20 minutes, heating off and cooling to room temperature (23-25 degrees Celsius) with stirring. The obtained nanoparticles had a particle size of 13 nm and a concentration of 10 nM. Transmission electron microscopy (TEM) showed that the obtained nanoparticles were spherical.
[0064] Since the sodium citrate-modified gold nanoparticles were unstable at high salt concentration...
Embodiment 2
[0065] Example 2: Preparation of single-stranded DNA probe-modified fluorescent quantum dots
[0066] Fluorescent quantum dots were prepared by one-step reaction (see: Q.B.Wang, D.K.Seo.JMater Sci., 2009, 44, 816; Q.B.Wang, Y.Liu, Y.Ke, H.Yan.Angew.Chem., Int. Ed., 2008, 47, 316; Q.B.Wang, N.Iancu, D.K.Seo.Chem.Mater., 2005,17:4762; and Q.B.Wang, Y.Xu, X.Zhao, Y.Chang, Y.Liu, L. Jiang, J. Sharma, D. K. Seo, H. Yan. J. Am. Chem. Soc., 2007, 129, 6380).
[0067] Specifically, MPA-modified CdSe / ZnS core / shell quantum dots were prepared as follows. Put 0.1 mmol P 2 S 5 , 0.5 g of 3-mercaptopropionic acid (MPA) and 0.3 ml of butylamine were mixed in 10 ml of N-methyl-2-pyrrolidone (NMP) and heated to dissolve. After cooling to room temperature, 0.01 g of oleylamine-modified CdSe core particles (provided by Beijing Zhongke Wuyuan Biotechnology Co., Ltd., with a fluorescence peak of 525 nm) were added, and then mixed with 0.05 g of ZnCl 2 solution mix. The mixed solution was he...
Embodiment 3
[0070] Example 3: Preparation of Fluorescent Quantum Dot / Nano Metal Particle Conjugates
[0071]In the QD-DNA solution from Example 2 with a QD concentration of 200 nM, an Au-DNA solution with a particle size of 13 nm from Example 1 with a concentration of Au of 8 nM was added, where QD:Au=Au=QD in the QD-Au structure was maintained 5:1, that is, 2 microliters of QD-DNA solution and 10 microliters of Au-DNA solution, and then add DNase (40 μM, 0.1 microliters) and the corresponding substrate DNA (40 μM) to the system. , 0.1 μl) and sodium chloride solution to a concentration of 0.15 mM sodium chloride, and annealed the hybridization overnight. The annealing curve for DNA hybridization was held at 80°C for 5 minutes and then slowly cooled to room temperature.
[0072] When the target is lead, the DNase is 5'-CAT CTC TTC TCC GAG CCGGTC GAA ATA GTG AGT-3' and the substrate DNA is 3'-GTG CTC AAC TGTGTA GAG AAG GrA TAT CAC TCA AGT GTC TAC TCA-5'.
[0073] When the target is copp...
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