Binding and purification of plasmid DNA using multi-layered carbon nanotubes |
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Authors: | Shakhmaeva Irina I Bulatov Emil R Bondar Oksana V Saifullina Diana V Culha Mustafa Rizvanov Albert A Abdullin Timur I |
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Institution: | a Kazan (Volga Region) Federal University, 18 Kremlyovskaya St., 420008 Kazan, Republic of Tatarstan, Russia b Department of Genetics and Bioengineering, Yeditepe University, 34755 Istanbul, Turkey c Core Research Laboratory, Kazan State Medical University, 49 Butlerova St., 420012 Kazan, Republic of Tatarstan, Russia |
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Abstract: | We propose a new method for the separation of nucleic acids using multi-layered carbon nanotubes (CNTs) as an adsorbent. According to agarose gel electrophoresis, oxidized water-stable CNTs adsorb certain forms of nucleic acids, such as high molecular weight RNA, chromosomal DNA, linear and denatured forms of plasmid DNA. However, CNTs do not adsorb supercoiled form of plasmid DNA. Nucleic acids bound to CNTs can be readily removed by centrifugation whereas supercoiled plasmid DNA remains in solution. Upon the addition of divalent metal ions supercoiled plasmid DNA forms relatively stable complexes with CNTs due to chelation. Thus, new details about association of nucleic acids with CNTs were revealed and stoichiometry of the complexes was estimated. Our results can be used for fine purification of supercoiled plasmid DNA for gene therapy applications as well as manipulation of nucleic acids for biosensor design. |
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Keywords: | Nucleic acid separation Plasmid DNA Gene therapy Adsorbents Carbon nanotubes Biosensors |
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