• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Current Protocols in Nucleic Acid Chemistry >NMR Spectroscopy of Large Functional RNAs: From Sample Preparation to Low-Gamma Detection
【24h】

NMR Spectroscopy of Large Functional RNAs: From Sample Preparation to Low-Gamma Detection

机译：核磁共振光谱学的大型功能rna:样品制备Low-Gamma检测

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

摘要

NMR spectroscopy is a potent method for the structural and biophysical characterization of RNAs. The application of NMR spectroscopy is restricted in RNA size and most often requires isotope-labeled or even selectively labeled RNAs. Additionally, newNMR pulse sequences, such as the heteronuclear-detected NMR experiments, are introduced. We herein provide detailed protocols for the preparation of isotope-labeled RNA for NMR spectroscopy via in vitro transcription. This protocol covers all steps, fromthe preparation of DNA template to the transcription of milligram RNA quantities. Moreover, we present a protocol for a chemo-enzymatic approach to introduce a single modified nucleotide at any position of any RNA. Regarding NMR methodology, we share protocols for the implementation of a suite of heteronuclear-detected NMR experiments including ~(13)C-detected experiments for ribose assignment and amino groups, the CN-spin filter heteronuclear single quantum coherence (HSQC) for imino groups and the ~(15)N-detected band-selective excitation short transient transverse-relaxation-optimized spectroscopy (BEST-TROSY) experiment.
机译：核磁共振光谱法是一种有效的方法结构和生物物理特性rna。限制在RNA的大小和最经常需要isotope-labeled甚至有选择性地标记rna。此外,newNMR脉冲序列,例如heteronuclear-detected核磁共振实验中,介绍了。isotope-labeled RNA的准备通过体外转录核磁共振光谱学。协议涵盖了所有步骤,从准备DNA的转录模板毫克RNA数量。chemo-enzymatic方法介绍单核苷酸在任何位置的任何修改RNA。协议实施一套heteronuclear-detected NMR实验包括~(13)为核糖C-detected实验任务和氨基酸组,CN-spin过滤器杂环的单量子相干(HSQC)亚氨基的团体和~ (15)N-detectedband-selective激发短瞬态transverse-relaxation-optimized光谱学(BEST-TROSY)实验。

著录项

相似文献

  • 外文文献
  • 中文文献
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