PCR is widely employed as the initial DNA amplification step for genetic testing. However, a key limitation of PCR-based methods is the inability to selectively amplify low levels of mutations in a wild-type background. As a result, downstream assays are limited in their ability to identify subtle genetic changes that can have a profound impact in clinical decision-making and outcome. Here we describe co-amplification at lower denaturation temperature PCR (COLD-PCR), a novel form of PCR that amplifies minority alleles selectively from mixtures of wild-type and mutation-containing sequences irrespective of the mutation type or position on the sequence. We replaced regular PCR with COLD-PCR before sequencing or genotyping assays to improve mutation detection sensitivity by up to 100-fold and identified new mutations in the genes encoding p53, KRAS and epidermal growth factor in heterogeneous cancer samples that had been missed by the currently used methods. For clinically relevant microdeletions, COLD-PCR enabled exclusive amplification and isolation of the mutants. COLD-PCR will transform the capabilities of PCR-based genetic testing, including applications in cancer, infectious diseases and prenatal identification of fetal alleles in maternal blood.

The combined effects of hydroxyl/amino functional groups of aromatics and surface O-containing groups of carbon nanotubes on adsorption were evaluated. When normalized for hydrophobicity, 2,4-dichlorophenol and 2-naphthol exhibited a greater adsorptive affinity to carbon nanotubes and graphite (a model adsorbent without the surface O functionality) than structurally similar 1,3-dichlorobenzene and naphthalene, respectively, and 1-naphthylamine exhibited a much greater adsorptive affinity than all other compounds. Results of the pH-dependency experiments further indicated that the hydroxy/amino functional groups of the adsorbates and the surface properties of the adsorbents played a combinational role in determining the significance of the nonhydrophobic adsorptive interactions. We propose that the strong adsorptive interaction between hydroxyl-substituted aromatics and carbon nanotubes/graphite was mainly due to the electron-donating effect of the hydroxyl group, which caused a strong electron-donor-acceptor (EDA) interaction between the adsorbates and the pi-electron-depleted regions on the graphene surfaces of carbon nanotubes or graphite. In addition to the EDA interaction, Lewis acid-base interaction was likely an extra important mechanism contributing to the strong adsorption of 1-naphthylamine, especially on the O-functionality-abundant carbon nanotubes. The findings of the present study might have significant implications for selective removal of environmental contaminants with carbon nanotubes.