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相关概念视频

Real Time RT-PCR02:57

Real Time RT-PCR

57.0K
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...
57.0K

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相关实验视频

Updated: Jun 17, 2025

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies
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在qRT-PCR分析中开发一个强大的和可泛化的算法"gQuant",用于准确的正常化基因选择.

Abhay Kumar Pathak1, Sukhad Kural2, Shweta Singh3

  • 1DST-CIMS, Institute of Science, Banaras Hindu University, Varanasi, India.

Scientific reports
|August 13, 2024
PubMed
概括

一个新的工具,gQuant,通过选择可靠的正常化基因用于qRT-PCR分析,提高基因表达量化的准确性. 这解决了使用microRNAs,长非编码RNAs和信使RNAs发现生物标记物的挑战.

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Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
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科学领域:

  • 生物标志物发现发现
  • 分子诊断学 分子诊断
  • 生物信息学是一种生物信息学.

背景情况:

  • 核酸生物标志物 (miRNAs, lncRNAs, mRNAs) 对于疾病诊断至关重要.
  • 定量逆转录PCR (qRT-PCR) 是生物标志物测量的标准.
  • 准确的qRT-PCR需要可靠的正常化基因,目前缺乏非编码RNA.

研究的目的:

  • 在qRT-PCR中开发用于正常化基因选择的强大工具.
  • 克服现有的规范化器选择工具中的统计局限性和可视化差异.
  • 为了提高生物标志物分析的基因表达量化的精度.

主要方法:

  • 开发了"gQuant",一种使用投票分类器结合多种统计方法的新工具.
  • 通过使用各种尿道外体miRNA数据集验证的gQuant.
  • 将gQuant的性能与现有的规范化器选择工具进行比较.

主要成果:

  • 与现有的工具相比,gQuant显示出优越的正常化基因排名.
  • 来自gQuant的排名显示了较低的标准偏差和协差.
  • 增强的内核密度估计值表明了改进的数据分布分析.
  • gQuant避免了有时由其他方法产生的偏斜排名.

结论:

  • gQuant显著改善了对qRT-PCR的正常化基因鉴定.
  • 该工具提高了在各种研究环境中基因表达量化的精度.
  • gQuant为分析qRT-PCR数据提供了一个平衡的解决方案,特别是对于非编码RNA生物标志物.
  • 该gQuant工具是公开可用的.