Abstract
An addressable electrode array was used for the production of acid at sufficient concentration to allow deprotection of the dimethoxytrityl (DMT) protecting group from an overlaying substrate bound to a porous reaction layer. Containment of the generated acid to an active electrode of 100 micron diameter was achieved by the presence of an organic base. This procedure was then used for the production of a DNA array, in which synthesis was directed by the electrochemical removal of the DMT group during synthesis. The product array was found to have a detection sensitivity to as low as 0.5 pM DNA in a complex background sample.
Highlights
Rapid developments in the field of DNA arrays have led to a number of methods for their in situ synthetic preparation, including photolithography using both fixed and programmable masks, ink jet printing of reagents, and electrochemical deprotection techniques.[1]
These techniques have led to a number of commercially available DNA microarray products, including the CombiMatrix product used in this report as well as systems produced by Affymetrix, Agilent, Nimblegen, and Febit
The overlaying porous reaction layer (PRL) provides a solid support for attachment of our product DNA, similar to the CPG used in the column of a commercial DNA synthesizer, while maintaining close proximity to the electrode
Summary
Rapid developments in the field of DNA arrays have led to a number of methods for their in situ synthetic preparation, including photolithography using both fixed and programmable masks, ink jet printing of reagents, and electrochemical deprotection techniques.[1] These techniques have led to a number of commercially available DNA microarray products, including the CombiMatrix product used in this report as well as systems produced by Affymetrix, Agilent, Nimblegen, and Febit. These systems rely on the standard phosphoramidite DNA synthesis procedure, as shown, with some modification of the deprotection or coupling procedures. The precise alignment and many steps of production in such systems have led us to consider the generation of acid by an electrochemical means for the deprotection of the DMT group during the array synthesis process
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