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相关概念视频

Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
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用基于核酸的测试来描述细胞自由转录和翻译动态.

Fernanda Piorino1,2, Chad Sundberg3, Elizabeth A Strychalski1

  • 1National Institute of Standard and Technology, Gaithersburg, Maryland 20899, United States.

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概括

这项研究引入了细胞自由表达 (CFE) 系统的新测量框架. 它为转录和翻译动态提供了可访问的定量指标,改善了CFE技术开发.

关键词:
没有细胞表达系统的细胞表达系统.标志性特征的描述.计量学 计量学 计量学 计量学转录 转录 是一种转录.翻译翻译翻译翻译翻译翻译

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Author Spotlight: Characterizing Novel Enzymes from Extremophiles and Common Pathogens to Understand DNA Repair and Replication
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科学领域:

  • 合成生物学 合成生物学
  • 生物技术是生物技术.
  • 分子生物学分子生物学

背景情况:

  • 目前的无细胞表达 (CFE) 系统表征严重依赖于有限的指标,如单个光蛋白测量.
  • 综合性表征具有挑战性,因为需要对涉及内源和外源成分的复杂反应进行时间过程测量.
  • 开发可预测和可重复的CFE技术需要更强大的性能指标.

研究的目的:

  • 开发和演示可访问的测量框架,用于描述CFE系统中的转录和翻译动态.
  • 提供有意义的定量指标,超越简单的光谱分析.
  • 为了使更可预测和可重复的CFE技术的发展.

主要方法:

  • 开发了一个新的测量框架来跟踪随时间推移的转录和翻译动态.
  • 该框架应用于各种 *Escherichia coli* CFE 系统,包括内部和商业提取物和复制系统.
  • 使用不同的核酸模板,并包括低性能系统来测试框架的灵敏度和确定性能指标.

主要成果:

  • 该研究成功计算了反应指标,以便在各种CFE系统中进行定量比较.
  • 测量框架在表征高性能和低性能CFE系统方面表现出了敏感性.
  • 阐明了表明系统性能的关键指标.

结论:

  • 开发的框架为全面的CFE系统表征提供了一种可访问的方法.
  • 这种方法补充了现有实践,并提供了对推进CFE技术至关重要的信息数据.
  • 该框架适用于CFE研发中的常规采用.