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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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DNA Microarrays02:34

DNA Microarrays

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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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相关实验视频

Updated: Jul 6, 2025

Measuring mRNA Levels Over Time During the Yeast S. cerevisiae Hypoxic Response
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在家进行定量mRNA表达测量.

Sonalisa Pandey1,2, Sara Safa McCoy2,3, Tsering Stobdan2

  • 1Shanvi, San Diego, CA, USA.

Scientific reports
|January 10, 2024
PubMed
概括

这项研究引入了一种基于光的快速反转录循环介导同热放大 (RT-LAMP) 方法,用于准确的mRNA测量. 这种易于使用的技术为昂贵的RT-PCR提供了可行的替代方案,使得诊断点的诊断成为可能.

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Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
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Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
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相关实验视频

Last Updated: Jul 6, 2025

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Measuring mRNA Levels Over Time During the Yeast S. cerevisiae Hypoxic Response

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科学领域:

  • 分子生物学分子生物学
  • 生物技术是生物技术.
  • 诊断 诊断 诊断 诊断

背景情况:

  • 反转录聚合酶连锁反应 (RT-PCR) 是mRNA测量的标准,但需要专门的设备和专业知识.
  • 循环介导同热放大 (LAMP) 是一种敏感的核酸检测方法,但其对RNA检测的灵敏度在历史上落后于RT-PCR.
  • 准确和可访问的mRNA量化对于各种生物和临床应用至关重要.

研究的目的:

  • 开发和验证基于光的反转录循环介导同热放大 (RT-LAMP) 协议,用于敏感和准确的mRNA量化.
  • 为了比较开发的RT-LAMP试验与已建立的RT-PCR方法的性能.
  • 证明RT-LAMP试验对不同样本类型的有用性及其对照料中心应用的潜力.

主要方法:

  • 一个基于光的新型RT-LAMP协议被开发用于测量CDX2mRNA表达.
  • 实施了一种新的原料设计策略,以最大限度地减少假阳性并提高量化准确性.
  • 该测试使用各种样本类型进行验证,包括cDNA,mRNA和直接组织样本.

主要成果:

  • 开发的RT-LAMP协议与CDX2表达的RT-PCR标准具有很高的相关性 (r=0.99,p<0.001).
  • 该试验在各种样本类型中提供了准确的量化,包括直接组织样本.
  • 整个测量过程,从样本到结果,大约在25分钟内完成,比RT-PCR (3小时以上) 快得多.

结论:

  • 基于光的RT-LAMP为mRNA测量提供了一个灵敏,准确和快速的RT-PCR替代方案.
  • 开发的协议和原料设计策略克服了LAMP灵敏度和特异性的先前限制.
  • 测试的简单性和速度,再加上一个便携式设备,便于在家中,户外和医院的mRNA表达分析.