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:: Volume 20, Issue 4 (2-2013) ::
Journal of Ilam University of Medical Sciences 2013, 20(4): 200-207 Back to browse issues page
Separation and Application of DNA Aptamer for Detection of Morphine
S Haidari kashl , Gh r Behroozi * , Ml Mosavi , Mj Rasaee , J Salimian , A Akbari
Abstract:   (13963 Views)
Introduction: With regard to the chara-cteristics of single-stranded oligonucle-otides, including their potential for implem-ental use in order to detect some key molec-ules, it was determined to gain experience in obtaining single-stranded DNA (ssDNA) having affinity for binding to morphine molecules. Materials, methods & Findings: An oligo-nucleotide library was prepared, consisting of an 80-nucleotide sequence with fixed length flanked by constant 5′ and 3′ ends, a 19-nucleotide sequence from 3′-end, a 21-nucleotide sequence from 5′-end that serve as primer, and a 40-nucleotide random seq-uence in the middle. Four types of nucleic acid and the 40-nucleotide sequence are mathematically equaled with 1015 DNA chains known as oligonucleotide pool. Af-ter amplification by PCR, the aptamers we-re exposed to the matrix-bound morphine molecules. After rinsing with a buffer and eliminating the chains, not bound to mor-phine molecule, they were separated using washing buffer, having potential to separate DNA chains from the target molecule. This act was repeated 15 times. Lastly, those primers were used in which one fluorescein molecule was placed at its 5′-end and flow cytometry results proved the efficiency of the chain in detection of target molecule (morphine). Discussion & Conclusion: Morphine mol-ecule was chosen due to its social and commercial importance for governments. For this reason, the separation of this nucl-eotide segment (oligonucleotide) was desi-gned by applying the experience of other researchers and some segments were event-ually obtained, which can be utilized inst-ead of monoclonal antibodies in detecting morphine in rapid kits after detection of sequences. Therefore, we enter to nanobi-ology and we will achieve applying olig-onucleotides in treatment, drug delivery into the target molecule, follow up the effect of the drug on cells, and finally help imaging.
Keywords: aptamer, morphine, SELEX
Full-Text [PDF 1102 kb]   (8442 Downloads)    
Type of Study: Research | Subject: exclusive bacteriology
Received: 2013/03/13 | Accepted: 2013/10/15 | Published: 2013/10/15
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haidari kashl S, behroozi G R, mosavi M, rasaee M, salimian J, akbari A. Separation and Application of DNA Aptamer for Detection of Morphine. J. Ilam Uni. Med. Sci. 2013; 20 (4) :200-207
URL: http://sjimu.medilam.ac.ir/article-1-910-en.html

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Volume 20, Issue 4 (2-2013) Back to browse issues page
مجله دانشگاه علوم پزشکی ایلام Journal of Ilam University of Medical Sciences
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