Aeromonas hydrophila aptamer, and screening method and application thereof
A technology of Aeromonas hydrophila and aptamer, which is applied in the field of Aeromonas hydrophila aptamer and its screening, can solve the problems of cumbersome detection process, high cost, time-consuming and the like
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[0023] 1. Synthesis of ssDNA library and primers
[0024] Referring to the method disclosed in the literature (7. Liu Fengwei, Lan Xiaopeng. In vitro screening of Pseudomonas aeruginosa aptamer research and preliminary application [D]. Fuzhou Fujian Medical University, 2007.), the random ssDNA library used for SELEX screening and primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.
[0025] The library used was: 5'-GGG AGC TCA GAA TAA ACG CTC AA-N35-TT CGA CAT GAG GCC CGG ATC-3'.
[0026] The primers used are:
[0027] Primer Ⅰ: 5′-GGG AGC TCA GAA TAA ACG CTC AA-3′;
[0028] Primer II: 5′-GAT CCG GGC CTC ATG TCG AA-3′;
[0029] Primer Ⅲ: 5′-Digoxigenin-GGG AGC TCA GAA TAA ACG CTC AA-3′;
[0030] Primer IV: 5'-biotin-GAT CCG GGC CTC ATG TCG AA-3'.
[0031] After the above primers were synthesized, they were diluted with TE buffer (pH 8.0) commonly used in molecular biology to a concentration of 10 μmol, and stored at -20°C for future use....
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