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相关概念视频

Evolutionary Relationships through Genome Comparisons02:54

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Genome comparison is one of the excellent ways to interpret the evolutionary relationships between organisms. The basic principle of genome comparison is that if two species share a common feature, it is likely encoded by the DNA sequence conserved between both species. The advent of genome sequencing technologies in the late 20th century enabled scientists to understand the concept of conservation of domains between species and helped them to deduce evolutionary relationships across diverse...
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Genome Annotation and Assembly03:36

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The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base pairs in many eukaryotic organisms. Genome assembly refers to the process of taking the DNA sequencing data and putting it all back together in a correct order to create a close representation of the original genome. This is followed by the identification of functional elements on the newly assembled genome, a process called genome annotation.
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相关实验视频

Updated: May 3, 2026

Sample Preparation and Analysis of RNASeq-based Gene Expression Data from Zebrafish
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基于转录基因组序列数据的基础上,选择和验证中的参考基因.

Wenna Fan1, Yaqi Shi2, Pengfei Shi2

  • 1Animal Science and Technology College, Henan University of Science and Technology, Luoyang, 471003, Henan, China. chou0516@163.com.

Scientific reports
|February 21, 2025
PubMed
概括

这项研究确定了在各种非生物压力下在中RT-qPCR的最佳内部参考基因. 像GAPDH和Actin这样的传统基因并不理想;对于精确的基因表达分析,建议使用特定的基因,如UBL-2a和Ms.33,066.

关键词:
阿尔法尔法 (Alfalfa) 是一种植物.参考基因是一种基因.选择 选择 选择 选择转录组序列的序列.

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Comprehensive Workflow for the Genome-wide Identification and Expression Meta-analysis of the ATL E3 Ubiquitin Ligase Gene Family in Grapevine
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科学领域:

  • 植物分子生物学 植物分子生物学
  • 基因组学和转录基因组学

背景情况:

  • 使用RT-qPCR进行定量基因表达分析,需要可靠的内部参考基因来准确规范化.
  • 传统的参考基因 (GAPDH,Actin) 可能在不同的实验条件下不稳定,特别是像 (Medicago sativa L.) 这样的植物中的非生物应激.

研究的目的:

  • 在五种不同的非生物压力条件下 (干旱,,高温,低温和酸性) 识别和验证中RT-qPCR最稳定的内部参考基因.
  • 评估常用的参考基因 (GAPDH,Actin) 和新型候选基因的适应性,以准确地规范的基因表达.

主要方法:

  • 候选参考基因从柳转录组数据 (162个RNA-seq数据集) 中进行了选择.
  • 在干旱,,高温和低温压力下使用RT-qPCR评估了10个候选基因的表达稳定性.
  • 用多种算法和软件评估基因稳定性,包括GeNorm,Normfinder,Bestkeeper,ΔCt方法和RefFinder.

主要成果:

  • 发现GAPDH和Actin在的各种非生物压力下不适合作为参考基因.
  • 确定了最佳的单基因参考基因:UBL-2a (),Ms.33,066 (干旱) 和Actin (高/低温).
  • 对 (MS.65,463和UBL-2a) 和干旱 (MS.65,463和UBL-2a) 的压力确定了最佳基因基因组合,对高温和低温建议进一步组合.

结论:

  • 这项研究为在非生物压力下在草基因表达研究中选择适当的参考基因提供了关键数据.
  • 建议使用新的参考基因和组合,比传统基因更准确地进行的RT-qPCR分析.
  • 在酸性压力下进一步验证最佳参考基因是有必要的.