A method for the detection of Salmonella by enzyme repair isothermal cycle amplification fluorescence method
A cyclic amplification and Salmonella technology, applied in the field of detection, can solve the problems of long detection cycle, low specificity and sensitivity, etc., and achieve the effect of fast detection speed, improved sensitivity and high specificity
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[0037] The preparation method of described fluorescent biosensor comprises the following steps:
[0038] (1) Add sterilized water, 10× buffer (buffer), 1 μL 20 μM aptamer chain and Trigger chain to the pre-prepared sterilized EP tube; shake for 30s, incubate at 95°C for 5min, slowly Cool to room temperature;
[0039] (2) Add 1 μL of target bacteria (concentrations of 10 6 cfu / mL, 10 5 cfu / mL, 10 4 cfu / mL, 10 3 cfu / mL, 10 2cfu / mL), UDG (1 μL), 10 μM HAP (1 μL), 10 μM Signal probe (1 μL), endonuclease IV (1 μL), Bst DNA polymerase (1 μL), and dNTP (1 μL) Shake for 30s, and incubate for 2 hours in a 37°C incubator;
[0040] (3 Dilute the above-mentioned reacted solution (10 μL) to 70 μL, and detect the fluorescence at 518nm with a fluorometer;
[0041] The excitation wavelength of the fluorescence instrument is set to 486nm, the emission wavelength is 518nm, the detection range is 450nm-530nm, and the change of the fluorescence signal is read to detect the target objec...
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