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使用RT-qPCR在正在发育的小鼠皮质中进行基因表达研究的适合参考基因的评估.

Ananya Uppalapati1, Timothy Wang1, Lena H Nguyen2

  • 1Department of Neuroscience, School of Behavioral and Brain Sciences, The University of Texas at Dallas, Richardson, TX, 75080, USA.

BMC neuroscience
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PubMed
概括

为了在发育中的小鼠皮质中进行准确的基因表达分析,建议使用B2m,Gapdh和Hprt作为实时定量PCR (RT-qPCR) 的参考基因. 这些基因在晚期皮质发生过程中表现稳定,这对于可靠的神经科学研究至关重要.

关键词:
大脑皮层的大脑皮层.基因表达 基因表达 基因表达家庭管理基因鼠标 鼠标是一个鼠标.神经发育 神经发育RT-qPCRR 是一个人.参考基因是指基因中的基因.

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科学领域:

  • 神经科学是一个神经科学.
  • 分子生物学分子生物学
  • 遗传学 是一个遗传学.

背景情况:

  • 实时定量PCR (RT-qPCR) 对于神经科学基因表达研究至关重要.
  • 稳定的参考基因对于准确的RT-qPCR相对量化至关重要.
  • 关于在发育中的小鼠皮质中参考基因稳定性的数据有限.

研究的目的:

  • 为了评估常用的参考基因在发育中的小鼠皮质的表达稳定性.
  • 在晚期皮质生成 (胚胎15日到产后0日) 期间识别RT-qPCR的最佳参考基因.

主要方法:

  • 评估了五种家政基因的稳定性:Actb,Gapdh,B2m,Rpl13a和Hprt.
  • 使用了五种计算算法:BestKeeper,geNorm,NormFinder,DeltaCt和RefFinder. 这三种算法都在使用中.
  • 从胚胎第15天到产后第0天的小鼠分析皮质组织.

主要成果:

  • B2m,Gapdh和Hprt显示出最稳定的表达.
  • B2m/Gapdh和B2m/Hprt的组合也显示出高稳定性.
  • Actb和Rpl13a表现出最不稳定的表达模式.

结论:

  • 在小鼠皮层发育研究中,B2m,Gapdh和Hprt被确定为适合RT-qPCR的参考基因.
  • 这些基因为从胚胎第15天到产后第0天的基因表达分析提供了可靠的正常化.
  • 这些发现支持在关键发育时期的神经科学研究中的准确相对量化.