A homogeneous rapid detection method for ochratoxin a based on nucleic acid chimeric dye fluorescence quenching
A technology of ochratoxin and detection method, which is applied in the field of fluorescence detection, can solve problems such as long detection time, locked operation, complicated scheme, etc., and achieves the effects of high specificity, simple and convenient operation, and good reproducibility
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[0020] The present invention will be described in detail below in conjunction with specific embodiments.
[0021] (1) Formation of fluorescent complexes between ochratoxin A nucleic acid aptamers and nucleic acid chimeric dyes
[0022] First, the ochratoxin A nucleic acid aptamer was pretreated: denatured at 95°C for 5 minutes, and cooled at 4°C for 5 minutes. Then take 50 μL of 10 mM Tris-HCl reaction buffer (pH 8.5), wherein the concentration of ochratoxin A nucleic acid aptamer is 50 nM, the dilution ratio of GeneFinder nucleic acid chimeric dye is 8:10000, and react at room temperature for 30 min to make nucleic acid chimeric The dye is fully combined with the ochratoxin A nucleic acid aptamer to form a fluorescent complex of the ochratoxin A nucleic acid aptamer and nucleic acid chimeric dye; the ochratoxin A nucleic acid aptamer is a single-stranded DNA probe with a sequence of 5' -CTTCGGGTGTGGGTGGCATTCCGGTAGGCTCTG-3'.
[0023] Screening and preparation of an ochratoxi...
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