English Extended Abstract: (16909 Views)
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
Article Type:
Systematic Review |
Subject:
Police Medicine Related Technologies Received: 2013/12/21 | Accepted: 2014/03/12 | Published: 2014/03/12