Preparation and detection methods of p-benzoquinone-mediated escherichia coli
A technology for Escherichia coli and detection methods, applied in the direction of microorganism-based methods, methods using microorganisms, biochemical equipment and methods, etc., can solve problems such as expensive, time-consuming, and time-consuming, and is suitable for large-scale promotion and production. The effect of simple preparation method and great application prospect
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Embodiment 1
[0043] Embodiment 1, microorganism cultivation
[0044] In step 1, the bacteria were placed in LB medium, and the bacterial culture was grown overnight at a culture temperature of 37° C., an oscillation frequency of 150 rpm, and a culture time of 12 hours.
[0045] In step 2, the bacterial culture solution obtained in step 1 was centrifuged at an oscillation frequency of 3600 rpm for 10 minutes and washed three times with phosphate buffered saline. Suspend the remaining bacterial pellet in phosphate-buffered saline for further use.
Embodiment 2
[0046] Embodiment 2, antibiotic resistant bacteria induction
[0047] Antibiotic selection: one of trimethoprim resistance, erythromycin resistance.
[0048] In step 1, wild-type E. coli (WT) was inoculated into a series of Erlenmeyer flasks with a double antibiotic concentration gradient, with a final volume of 100 mL. Select the initial concentration of the double antibiotic concentration gradient to be 50% MIC (minimum inhibitory concentration), and each flask is shaken at a temperature of 37° C. and a shaking frequency of 150 rpm;
[0049] Step 2, measure the OD of each flask after step 2 600 Value (absorbance value of solution at 600nm wavelength);
[0050] Step 3, as the bacteria grow, the strain with the highest antibiotic concentration grown by the bacteria is propagated into another round of antibiotic resistance incubation with a higher antibiotic concentration gradient;
[0051] Step 4, after 5 rounds of selection, collect low antibiotic resistance Escherichia co...
Embodiment 3
[0055] Embodiment 3, the response of different concentrations p-benzoquinone in electrochemical method
[0056] Step 1, equipped with various concentrations of BQ (1, 2, 3, 4, 5, 6, 7, 8, 9 and 10mM) and E. coli (1.0×10 9 CFU / mL);
[0057] Step 2, get 3mL of various concentrations of BQ and 3mL Escherichia coli (1.0 × 10 9 CFU / mL) were incubated in centrifuge tubes for 1 hour.
[0058] In step 3, the sample obtained in step 2 was centrifuged at 3600 rpm for 10 minutes. The supernatant obtained from the sample was used for electrochemical detection.
[0059] The traditional three-electrode system is used to monitor the sample, and the CV curve of the interface is obtained by drawing the relationship between the current signal and the potential to obtain the cyclic voltammetry (CV) curve. The following conclusions can be drawn:
[0060] 1. With the increase of BQ concentration, the redox peak of CV curve is enhanced.
[0061] 2. Under the condition of the same volume of pho...
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