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Proceedings Paper

Porous silicon microparticles as an alternative support for solid phase DNA synthesis
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Paper Abstract

Current methods to produce short DNA strands (oligonucleotides) involve the stepwise coupling of phosphoramidites onto a solid support, typically controlled pore glass. The full-length oligonucleotide is then cleaved from the solid support using a suitable aqueous or organic base and the oligonucleotide is subsequently separated from the spent support. This final step, albeit seemingly easy, invariably leads to increased production costs due to increased synthesis time and reduced yields. This paper describes the preparation of a dissolvable support for DNA synthesis based on porous silicon (pSi). Initially it was thought that the pSi support would undergo dissolution by hydrolysis upon cleavage of the freshly synthesised oligonucleotide strands with ammonium hydroxide. The ability to dissolve the solid support after completion of the synthesis cycle would eliminate the separation step required in current DNA synthesis protocols, leading to simpler and faster synthesis as well as increased yields, however it was found that the functionalisation of the pSi imparted a stability that impeded the dissolution. This strategy may also find applications for drug delivery where the controlled release of carrier-immobilised short antisense DNA is desired. The approach taken involves the fabrication of porous silicon (pSi) microparticles and films. Subsequently, the pSi is oxidised and functionalised with a dimethoxytrityl protected propanediol to facilitate the stepwise solid phase synthesis of DNA oligonucleotides. The functionalisation of the pSi is monitored by diffuse reflectance infrared spectroscopy and the successful trityl labelling of the pSi is detected by UV-Vis spectroscopy after release of the dimethoxytrityl cation in the presence of trichloroacetic acid (TCA). Oligonucleotide yields can be quantified by UV-Vis spectroscopy.

Paper Details

Date Published: 19 January 2006
PDF: 10 pages
Proc. SPIE 6036, BioMEMS and Nanotechnology II, 60361W (19 January 2006); doi: 10.1117/12.659972
Show Author Affiliations
Steven McInnes, Flinders Univ. (Australia)
Sean Graney, Flinders Univ. (Australia)
Yit-lung Khung, Flinders Univ. (Australia)
Nicolas H. Voelcker, Flinders Univ. (Australia)

Published in SPIE Proceedings Vol. 6036:
BioMEMS and Nanotechnology II
Dan V. Nicolau, Editor(s)

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