DNA adsorption to and elution from silica surfaces: influence of amino acid buffers.
Solid phase extraction and purification of DNA from complex samples typically requires chaotropic salts that can inhibit downstream polymerase amplification if carried into the elution buffer. Amino acid buffers may serve as a more compatible alternative for modulating the interaction between DNA and silica surfaces. We characterized DNA binding to silica surfaces, facilitated by representative amino acid buffers, and the subsequent elution of DNA from the silica surfaces. Through bulk depletion experiments, we found that more DNA adsorbs to silica particles out of positively compared to negatively charged amino acid buffers. Additionally, the type of the silica surface greatly influences the amount of DNA adsorbed and the final elution yield. Quartz crystal microbalance experiments with dissipation monitoring (QCM-D) revealed multiphasic DNA adsorption out of stronger adsorbing conditions such as arginine, glycine, and glutamine, with DNA more rigidly bound during the early stages of the adsorption process. The DNA film adsorbed out of glutamate was more flexible and uniform throughout the adsorption process. QCM-D characterization of DNA elution from the silica surface indicates an uptake in water mass during the initial stage of DNA elution for the stronger adsorbing conditions, which suggests that for these conditions the DNA film is partly dehydrated during the prior adsorption process. Overall, several positively charged and polar neutral amino acid buffers show promise as an alternative to methods based on chaotropic salts for solid phase DNA extraction.
Keck Graduate Institute of Applied Life Sciences , 535 Watson Drive, Claremont, California 91711, United States.
Peter E
PE
Vandeventer
Jorge
J
Mejia
Ali
A
Nadim
Malkiat S
MS
Johal
Angelika
A
Niemz
D000327:Adsorption; D000596:Amino Acids / Q000737:chemistry*; D002021:Buffers; D004247:DNA / Q000737:chemistry*; D058525:Quartz Crystal Microbalance Techniques; D012822:Silicon Dioxide / Q000737:chemistry*; D052616:Solid Phase Extraction / Q000379:methods*; D013499:Surface Properties
D000596:Amino Acids; D002021:Buffers; D012822:Silicon Dioxide; D004247:DNA