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

Abstract

Impact of dispersion quality of NH 2 -MWCNTs (13-18 nm in diameter with a length between 1 and 12 lm, [99 % purity) in the amplification efficiency of a random DNA oligonucleotide library (96 bp) was investigated. Amplification yield in the presence of non-filtered NH 2 -MWCNT dispersion, filtered NH 2 -MWCNT dispersion and surfaceattached NH 2 -MWCNTs was explored, and physical interactions between NH 2 -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 NH 2 -MWCNT dispersion which was sonicated, centrifuged and filtered, enhanced the total PCR efficiency up to 70 % while the presence of NH 2 -MWCNT only centrifuged after sonication, inhibited the reaction significantly at similar concentrations. Furthermore, the NH 2 -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 NH 2 and non-filtered NH 2 -MWCNT water dispersions, which justifies the inhibition of the amplification by the non-filtered NH 2 -MWCNTs containing big agglomerates and bundles. Finally, we demonstrated that major PCR components adsorb onto the NH 2 -MWCNTs with diverse affinities, and maintain their functions after adsorption, which provides a good framework to further develop tunable NH 2 -MWCNT-carriers to be utilized in various nanobiotechnology and material science applications.Keywords Polymerase chain reaction Á Nano PCR Á Amine functionalized multiwall carbon nanotubes Á Random DNA oligonucleotide library Á Magnetic beads Guest Editors: Mustafa Culha, Rawil F. Fakhrullin, Ratnesh Lal This article is part of the topical collection on Nanobiotechnology Electronic supplementary material The online version of this article (

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