DNA three-way junction-silver cluster ratio type fluorescence sensor and method for detecting chloramphenicol
A fluorescent sensor and ratiometric technology, applied in the field of analysis and detection, can solve the problems of inaccuracy and inconvenience of residual chloramphenicol detection, and achieve the effects of obvious visual perception, improved dynamic response range, and stable fluorescence properties
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Embodiment 1
[0044] Example 1 Construction of DNA three-way junction-silver cluster ratiometric fluorescent sensor
[0045] (1) Connection of DNA three-way junction:
[0046] Put the three DNA samples at a molar ratio of 1:1:1 (Sequence 1: 5'-CCCCCTTAATCCCCCTTTTTTT-3', Sequence 2: 5'-AAAAAAACCCCCTAATTCCCCC-3', Sequence 3: 5'-ACTTCAGTGAGTTGTCCCACGGTCGGCGAGTCGGTGGTAG-3') in a centrifuge tube Hybridization was carried out in the system so that the final concentration of different DNAs in the system was the same, which was 100 μmol / L. Put the centrifuge tube containing the hybridized DNA sample into a water bath for annealing at 95°C for 5 min, then take out the centrifuge tube and slowly cool down (about 1 min to cool down). 1° C.), cooled to room temperature to obtain a DNA tee junction sample solution with a concentration of 33.33 μmol / L, and set aside.
[0047] (2) Preparation of AgNCs:
[0048] Take 75 μL of the DNA tee junction sample solution obtained in (1) in 805 μL ultrapure water ...
Embodiment 2
[0050] A series of known concentrations of chloramphenicol standard samples (0.01nmol / L, 0.05nmol / L, 0.1nmol / L, 0.2nmol / L, 0.3nmol / L, 0.4nmol / L, 0.5nmol / L, 1nmol / L L, 2nmol / L, 5nmol / L, 10nmol / L, 20nmol / L, 50nmol / L, 80nmol / L, 100nmol / L) were added to the DNA three-way junction-silver cluster ratiometric fluorescent sensor obtained in Example 1 In the dispersion liquid, the fluorescence intensity F obtained when the concentration is 0 0 , and the fluorescence intensity F when the concentration is not 0, calculate the relative fluorescence intensity I corresponding to each concentration F =(F 0 -F) / F 0 , using the relative fluorescence intensity and chloramphenicol concentration to construct a linear detection model (such as figure 2 shown). The regression equation is Y=1.0543X+0.0867, linear correlation (R 2 =0.9887), the linear range is within 0.01-0.5nmol / L, and the lowest detection limit is 9.8pmol / L.
[0051] Such as figure 1 Schematic diagram of the detection princi...
Embodiment 3
[0056] Example 3 Standard addition recovery verification
[0057] In order to examine the accuracy and reliability of this method, recovery experiments were carried out on the concentration of chloramphenicol in the linear range (0.01nmol / L, 0.1nmol / L, 0.5nmol / L) in the treated milk. The recovery rate was 90.37%-106.72%. The results showed that the method had good accuracy and could be detected in complex raw milk.
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