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Abstract
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BACKGROUND: The aim of this work is to achieve a DNA sequence of the aptamer that specifically able to detect of the trinitrotoluene (TNT).
MATERIALS AND METHODS: Amplification of nucleotide fragments (PCR) was performed using special primers and a library of random nucleotides (two determined sequences of 19 and 21 nucleotides for connecting to primers and 78 randomized nucleotides in the center). To SELEX (Systematic Evolution of Ligands by Exponential Enrichment) study using magnetic nanoparticles and the EDC reagent, TNT as binds to albumin (TNP-BSA) was immobilized. After initial screening, PCR were repeated on isolated pieces of nanoparticles using digoxigenin (DIG)-labeled nucleotides. In order to evaluate the aptamer function, using ELONA technique (Enzyme Linked Oligonucleotide Assay) and a specific anti-DIG antibody which is conjugated to peroxidase, the performance of aptamer for TNT detection was studied. Finally, this sequence is cloned into pBluescript plasmid and sequenced.
Results and Conclusion: The cloned aptamer has good efficiency for detection of TNT and could be used as aptasensor for detection of TNT in future studies
Language: fa |