2012
DOI: 10.1021/jp3017776
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Multiphasic DNA Adsorption to Silica Surfaces under Varying Buffer, pH, and Ionic Strength Conditions

Abstract: Reversible interactions between DNA and silica are utilized in the solid phase extraction and purification of DNA from complex samples. Chaotropic salts commonly drive DNA binding to silica, but inhibit DNA polymerase amplification. We studied DNA adsorption to silica using conditions with or without chaotropic salts through bulk depletion and Quartz Crystal Microbalance (QCM) experiments. While more DNA adsorbed to silica using chaotropic salts, certain buffer conditions without chaotropic salts yielded a sim… Show more

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Cited by 107 publications
(119 citation statements)
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“…[19]. Microscopic acid-fast staining direct and concentrated smears are prepared from sputum samples.…”
Section: Collection and Processing Of Pulmonary Sputum Samplementioning
confidence: 99%
“…[19]. Microscopic acid-fast staining direct and concentrated smears are prepared from sputum samples.…”
Section: Collection and Processing Of Pulmonary Sputum Samplementioning
confidence: 99%
“…This leads to electrostatic repulsion, due to the negatively charged silica surface and the negative charge of the DNA phosphate backbone. In order to compensate the electrostatic repulsion and promote the attachment of DNA, the chaotropic agent guanidinium thiocyanate was used in the binding buffer (Vandeventer et al, 2012). Thus, it is expected that both dsDNA tagged amplicons and ssDNA tagging primers will be attached on the silica magnetic particles surface, however, in higher extension for dsDNA tagged amplicons because of their higher negative charge density compared with ssDNA primers.…”
Section: Confocal Microscopy Of the Tagged Amplicon Immobilised On Simentioning
confidence: 99%
“…In a model proposed by Melzak et al, binding is driven first by the electrostatic shielding of the negative charges of the phosphate backbone of DNA and silica, second by the disruption of ordered water molecules that solvate each of the binding partners by chaotropic salts and organic solvents (entropy), and third by the formation of intermolecular hydrogen bonds (25). Subsequent studies have elaborated specific aspects of the model, such as the roles of buffer, pH, and ionic strength in the absence of chaotropic salts (26). The equilibrium between free double-stranded DNA and silica-bound DNA is sensitive to the concentrations of several reactants: protons (pH), salts (ionic strength), chaotrope (guanidine hydrochloride), organic solvent (alcohol), DNA, competing contaminants (including undesired nucleic acids), and silica (dimensions of column).…”
Section: Stick-less Silicamentioning
confidence: 99%