Dispersion quality of amine functionalized multiwall carbon nanotubes plays critical roles in polymerase chain reaction enhancement

Yüce, Meral and Budak, Hikmet (2014) Dispersion quality of amine functionalized multiwall carbon nanotubes plays critical roles in polymerase chain reaction enhancement. Journal of Nanoparticle Research, 16 (12). ISSN 1388-0764 (Print) 1572-896X (Online)

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Abstract

Impact of dispersion quality of NH2-MWCNTs (13-18 nm in diameter with a length between 1 and 12 mu m, >99 % purity) in the amplification efficiency of a random DNA oligonucleotide library (96 bp) was investigated. Amplification yield in the presence of non-filtered NH2-MWCNT dispersion, filtered NH2-MWCNT dispersion and surface-attached NH2-MWCNTs was explored, and physical interactions between NH2-MWCNTs and major PCR reagents including DNA template, wild type Taq DNA polymerase enzyme and primers were determined using high resolution polyacrylamide gel electrophoresis, dynamic light scattering, UV-Vis-NIR spectroscopy and scanning electron microscopy techniques. The results revealed that presence of NH2-MWCNT dispersion which was sonicated, centrifuged and filtered, enhanced the total PCR efficiency up to 70 % while the presence of NH2-MWCNT only centrifuged after sonication, inhibited the reaction significantly at similar concentrations. Furthermore, the NH2-MWCNTs coupled covalently onto magnetic microspheres, contributed for the specificity enhancement whilst decreasing the amplification efficiency by 30 % at the maximum concentration, which suggests a removable enhancement system for sensitive applications. On the other hand, the relative hydrodynamic size distribution measurements displayed a clear difference between the filtered NH2 and non-filtered NH2-MWCNT water dispersions, which justifies the inhibition of the amplification by the non-filtered NH2-MWCNTs containing big agglomerates and bundles. Finally, we demonstrated that major PCR components adsorb onto the NH2-MWCNTs with diverse affinities, and maintain their functions after adsorption, which provides a good framework to further develop tunable NH2-MWCNT-carriers to be utilized in various nanobiotechnology and material science applications.
Item Type: Article
Additional Information: Article Number: 2768
Uncontrolled Keywords: Polymerase chain reaction; Nano PCR; Amine functionalized multiwall carbon nanotubes; Random DNA oligonucleotide library; Magnetic beads
Subjects: Q Science > Q Science (General)
Divisions: Faculty of Engineering and Natural Sciences > Academic programs > Biological Sciences & Bio Eng.
Sabancı University Nanotechnology Research and Application Center
Faculty of Engineering and Natural Sciences
Depositing User: Meral Yüce
Date Deposited: 20 Nov 2015 11:13
Last Modified: 23 Aug 2019 11:27
URI: https://research.sabanciuniv.edu/id/eprint/27574

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