...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Analytical Biochemistry: An International Journal of Analytical and Preparative Methods >Examination of real-time polymerase chain reaction methods for the detection and quantification of modified siRNA
【24h】

Examination of real-time polymerase chain reaction methods for the detection and quantification of modified siRNA

机译：实时检测聚合酶链式反应的实时聚合酶链反应方法

获取原文
获取原文并翻译 | 示例
   

获取外文期刊封面封底 >>

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

摘要

With the ongoing efforts to develop siRNA-based therapeutics, there is a need for high-throughput detection and quantification of siRNA. Here we report the application of four reverse-transcriptase RT-PCR-based assays for the detection of 2'-deoxy-2'-fluoro and 2'-O-methyl-modified therapeutic siRNA in mouse plasma and tissue. These assays take advantage of the dynamic range, sensitivity, specificity, and high-throughput potential found in PCR assays. Three of these assays require design and optimization of primers and/or probes specific to the siRNA while the fourth utilizes a "universal" TaqMan probe that is independent of the siRNA sequence, thereby reducing method development time and cost. For the universal assay the range of detection in mouse plasma was 500 to 5e(-5) pg/mu l for four of five model Luciferase sequences tested. We found that the universal RT-PCR assay had comparable or better sensitivity and specificity than the other three assays. The universal design provides a rapid, sensitive, and specific assay with minimal method development time that will be well suited for high-throughput analysis of various siRNA sequences. (C) 2008 Elsevier Inc. All rights reserved.
机译：随着开发基于siRNA的治疗剂的不断努力，需要高通量的siRNA检测和定量。在这里，我们报告了基于四种逆转录酶RT-PCR的检测在小鼠血浆和组织中2'-脱氧-2'-氟和2'-O-甲基修饰的治疗性siRNA的检测的应用。这些测定利用了PCR测定中发现的动态范围，灵敏度，特异性和高通量潜力。这些测定法中的三种需要设计和优化siRNA特异性的引物和/或探针，而第四种则采用独立于siRNA序列的“通用” TaqMan探针，从而减少了方法开发的时间和成本。对于通用分析，对于五个测试的模型荧光素酶序列中的四个，小鼠血浆中的检测范围为500至5e（-5）pg /μl。我们发现，通用RT-PCR测定法具有比其他三种测定法相当或更好的敏感性和特异性。通用设计以最少的方法开发时间提供了快速，灵敏和特异的测定方法，非常适合各种siRNA序列的高通量分析。 （C）2008 Elsevier Inc.保留所有权利。

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