• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文会议>Conference on BioMEMS and Nanotechnology >Porous Silicon Microparticles as an Alternative Support for Solid Phase DNA Synthesis
【24h】

Porous Silicon Microparticles as an Alternative Support for Solid Phase DNA Synthesis

机译：多孔硅微粒作为固相DNA合成的替代载体

获取原文
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

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.
机译：产生短DNA链（寡核苷酸）的目前的方法涉及磷光酰胺的逐步偶联在固体载体上，通常控制孔玻璃。然后使用合适的含水或有机碱从固体载体中切割全长寡核苷酸，并且随后将寡核苷酸与支撑载体分离。最后一步，虽然看似容易，因此由于合成时间和产量降低而导致生产成本增加。本文描述了基于多孔硅（PSI）的DNA合成的可溶解载体的制备。最初据认为，PSI支持将通过水解在用氢氧化铵的新合成的寡核苷酸链中的水解时进行溶解。溶解合成循环后的固体载体的能力将消除当前DNA合成方案中所需的分离步骤，导致更简单和更快的合成以及增加的产率，然而发现PSI的功能赋予稳定性阻碍了溶解。该策略还可以找到药物递送的应用，其中需要载体固定化的短反义DNA的受控释放。采取的方法包括制造多孔硅（PSI）微粒和薄膜。随后，用二甲氧基特拉基受保护的丙二醇氧化并官能化，以促进DNA寡核苷酸的逐步固相合成。通过漫反射红外光谱监测PSI的官能化，并通过UV-Vis光谱在释放三甲氧基乙酸（TCA）存在下的二甲氧基酮阳离子后，检测PSI的成功Trityl标记。可以通过UV-Vis光谱量来量化寡核苷酸产率。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号