An oligonucleotide aptamer that specifically recognizes patulin
A patulin and oligonucleotide technology, which is applied in the preparation of test samples, DNA/RNA fragments, material inspection products, etc., can solve the problems of high detection cost, high precision, and low detection limit, etc. Achieve chemical stability, high specificity and affinity, and convenient results
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[0013] The following is a further description of screening nucleic acid aptamers that specifically bind to patulin by SELEX technology.
[0014] 1. Synthesize random single-stranded DNA library and primers (synthesized by IDT)
[0015] Random single-stranded DNA (ssDNA) library: 5′-CAGCTCAGAAGCTTGATCCT(N40)GACTCGAAGTCGTGCATCTG-3′, a random ssDNA library with a length of 80 nt was constructed, with fixed primer sequences at both ends and a random sequence of 40 bases in the middle, the library capacity for 10 14 Above; upstream primer Ⅰ: 5′-CAGCTCAGAAGCTTGATCCT-3′; downstream primer Ⅱ: 5′-CAGATGCACGACTTCGAGTC-3′, the random ssDNA library and the two primers were prepared into 100 μmol / L stock solution with TE buffer and stored at -20°C spare.
[0016] 2. Preparation and purification of ssDNA
[0017] The reaction system is: template DNA 5 μL, FAM-labeled upstream primer 1 μL (20 mM), phosphorylated downstream primer 0.5 μL (20 mM), dNTPmix (25 mM) 0.5 μL, 10×PCR amplificatio...
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