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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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Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
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通过使用基于qPCR的FOXO目标基因表达分析来测量FOXO活动.

Lucia Jimenez1, Inês Grenho2,3, Bruno Santos2,3

  • 1Sols-Morreale Biomedical Research Institute (IIBM), Spanish National Research Council (CSIC), Universidad Autónoma de Madrid (UAM), Madrid, Spain.

Methods in molecular biology (Clifton, N.J.)
|November 20, 2024
PubMed
概括

测量FOXO转录因子 (TF) 激活需要评估由于组织特异性表达的多个目标基因. 本研究介绍了一种使用定量逆转录聚合酶连锁反应 (qRT-PCR) 的方法,以准确测量FOXO TF活性.

关键词:
福克斯OFOXO 在线播放在量化方面,它是有数量的.目标 目标 目标 目标转录因子是一种转录因子.qRT-PCR 是一个很好的方法.

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科学领域:

  • 分子生物学分子生物学
  • 遗传学 是一个遗传学.
  • 细胞生物学 细胞生物学

背景情况:

  • FOXO转录因子 (TFs) 是哺乳动物的关键调节者,通过它们的分叉 (FKH) DNA结合域来识别.
  • 哺乳动物FOXO TFs (FOXO1,FOXO3,FOXO4,FOXO6) 是C. elegans daf-16的同类物,并与DAF-16家族蛋白质结合部位 (DBE) 结合.
  • FOXO TF活性至关重要,但由于目标基因的组织特异性表达,其评估是复杂的.

研究的目的:

  • 为了应对普遍测量FOXO转录因子活性的挑战.
  • 提出一种可靠的方法来评估在不同细胞环境中的FOXO激活.
  • 介绍一组目标基因组,这些基因集体表示FOXO活动.

主要方法:

  • 使用定量逆转录聚合酶链反应 (qRT-PCR) 来测量基因表达.
  • 开发一个协议,同时评估多个FOXO标基因.
  • 分析选定FOXO目标基因的集体表达模式.

主要成果:

  • 确定了一组FOXO目标基因,这些基因一起分析时,可以准确地测量FOXO激活.
  • 证明没有单一的FOXO点基因是FOXO活动的通用指标.
  • 通过多重基因表达分析建立了一个可靠量化FOXO TF活动的框架.

结论:

  • 为了准确测量FOXO转录因子活性,需要对特定目标基因进行集体评估.
  • 提出的qRT-PCR协议提供了一种可靠的方法来测量FOXO激活.
  • 这种方法克服了监测FOXO TFs中的组织特异性基因表达模式的局限性.