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Optimized in situ DNA synthesis on patterned glass

Published online by Cambridge University Press:  31 January 2011

Ishtiaq Saaem ,
Kuosheng Ma  and
Jingdong Tian
Ishtiaq Saaem
Affiliation:
ies2@duke.edu, Duke University, Biomedical Engineering, Durham, North Carolina, United States
Kuosheng Ma
Affiliation:
ucibiomems@gmail.com
Jingdong Tian
Affiliation:
jtian@duke.edu, Duke University, Biomedical Engineering, Durham, North Carolina, United States
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Abstract

This paper describes studies of patterned arrays on glass surfaces and their use as spatially separated reactors for in situ synthesis of DNA using an inkjet synthesizer. Photolithographic methods were employed to fabricate arrays composed of homogenous circular features containing a hydroxyl-terminated silane coupled to the surface of the glass via a siloxane bond. Features are embedded within a background matrix composed of a fluorosilane attached to the glass. Due to the differential wettability of the two silanes, whereby the hydroxyl-terminated silane and fluorosilane are hydrophilic and hydrophobic respectively because of their head groups, the patterned circular features are able to constrain liquid within a defined site. The silanization result was analyzed using X-ray photoelectron spectroscopy (XPS) to optimize silanization time and solvent. Synthesis was then performed using a custom-built inkjet system using phosphoramidite chemistry. Base-by-base analysis using fluorescent labeling showed consistent coupling efficiency on synthesis of a 50-mer homopolymer.

Keywords

biological synthesis (chemical reaction)ink-jet printingsurface chemistry
Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 1236: Symposium SS – Biosurfaces and Biointerfaces , 2009 , 1236-SS08-24
Copyright
Copyright © Materials Research Society 2010

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References

1 Patwardhan, R.P. et al. , High-resolution analysis of DNA regulatory elements by synthetic saturation mutagenesis. Nat Biotech, 2009. 27(12): p. 1173–1175.Google Scholar
2 Hughes, T.R. et al. , Expression profiling using microarrays fabricated by an ink-jet oligonucleotide synthesizer. Nat Biotech, 2001. 19(4): p. 342–347.Google Scholar
3 LeProust, E. et al. , Characterization of oligodeoxyribonucleotide synthesis on glass plates. Nucleic Acids Research, 2001. 29(10): p. 2171–2180.Google Scholar
4 Lausted, C. et al. , POSaM: a fast, flexible, open-source, inkjet oligonucleotide synthesizer and microarrayer. Genome Biology, 2004. 5(8): p.Google Scholar