• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Chemistry: A European journal >Tuning Single Nucleotide Discrimination in Polymerase Chain Reactions (PCRs):Synthesis of Primer Probes Bearing Polar 4'-C-Modifications and Their Application in Allele-Specific PCR
【24h】

Tuning Single Nucleotide Discrimination in Polymerase Chain Reactions (PCRs):Synthesis of Primer Probes Bearing Polar 4'-C-Modifications and Their Application in Allele-Specific PCR

机译：调整聚合酶链反应（PCR）中的单核苷酸歧视：带有极性4'-C-修饰的引物探针的合成及其在等位基因特异性PCR中的应用

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

There are many methods available for the detection of nucleo-tide variations in genetic material.Most of these methods are applied after amplification of the target genome sequence by the polymerase chain reaction (PCR).Many efforts are currently underway to develop techniques that can detect single nucleotide variations in genes either by means of,or without the need for,PCR.Allele-specific PCR (asPCR),which reports nucleotide variations based on either the presence or absence of a PCR-am-plified DNA product,has the potential to combine target amplification and analysis in one single step.The principle of asPCR is based on the formation of matched or mismatched primer-target complexes by using allele-specif-ic primer probes.PCR amplification by a DNA polymerase from matched 3'-primer termini proceeds,whereas a mismatch should obviate amplification.Given the recent advancements in realtime PCR,this technique should,in principle,allow single nucleotide variations to be detected online.However,this method is hampered by low selectivity,which necessitates tedious and costly manipulations.Recently,we reported that the selectivity of asPCR can be significantly increased through the employment of chemically modified primer probes.Here we report further significant advances in this area.We describe the synthesis of various primer probes that bear polar 4'-C-modified nucleotide residues at their 3' termini,and their evaluation in realtime asPCR.We found that primer probes bearing a 4'-C-methoxymethy-lene modification have superior properties in the discrimination of single nucleotide variations by PCR.
机译：检测遗传物质中核苷酸变化的方法有很多，其中大多数是在通过聚合酶链反应（PCR）扩增靶基因组序列后应用的，目前正在进行许多努力来开发可检测基因的方法。通过或不需要PCR的基因中的单个核苷酸变异。等位基因特异性PCR（asPCR）具有潜在的潜力，可根据是否存在PCR扩增的DNA产物报告核苷酸变异。 asPCR的原理是通过使用等位基因特异性引物探针形成匹配的或错配的引物-靶标复合物，基于DNA聚合酶从匹配的3'引物进行PCR扩增考虑到实时PCR的最新进展，从原理上讲，该技术应允许在线检测单核苷酸变异。H然而，该方法由于选择性低而受阻，这需要乏味且昂贵的操作。最近，我们报道了通过使用化学修饰的引物探针可以显着提高asPCR的选择性。在此我们报告该领域的进一步重要进展。描述了在3'末端带有极性4'-C修饰核苷酸残基的各种引物探针的合成，并通过PCR进行了实时评估。我们发现带有4'-C-甲氧基甲基镜片修饰的引物探针具有优越的性能PCR鉴别单核苷酸变异。

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