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相关概念视频

Real Time RT-PCR02:57

Real Time RT-PCR

64.6K
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...
64.6K

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相关实验视频

Updated: Jan 15, 2026

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
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基因表达分析的初学者一步步指南使用QIAcuity数字PCR基因表达分析.

Jonathan Weiss1, Gerald Schock2, Afif Abdel Nour2

  • 1QIAGEN GmbH, Hilden, Germany. Jonathan.Weiss@qiagen.com.

Methods in molecular biology (Clifton, N.J.)
|October 10, 2025
PubMed
概括

数字PCR (dPCR) 通过分割反应来提供精确的基因表达分析,以获得绝对量化. 这种方法克服了定量PCR (qPCR) 的局限性,为新的研究途径提供了更好的准确性和抑制剂耐药性.

关键词:
数字PCR (dPCR) 是一个数字PCR技术.基因表达 (Gex) 的基因表达多重复合是一种多重复合.QIA 电流量 电流量在RNA定量化过程中.

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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.
  • 生物技术是生物技术.

背景情况:

  • 基因表达 (GEX) 是一种基本的生物过程,涉及信使核糖核酸 (mRNA),对细胞功能至关重要.
  • 严格监管GEX对于发展和环境反应至关重要;异常的GEX与疾病有关.
  • 定量PCR (qPCR) 是一种常见的方法,但在效率,抑制剂和低丰度标方面存在局限性.

研究的目的:

  • 突出数字PCR (dPCR) 对基因表达分析的优势.
  • 在精度,准确性和限制方面,将dPCR与定量PCR (qPCR) 进行比较.
  • 展示dPCR如何使以前无法获得的基因表达研究成为可能.

主要方法:

  • 数字PCR (dPCR) 使用反应分区来进行绝对核酸量化,没有标准曲线.
  • 对dPCR和qPCR性能进行比较,重点关注准确性,可变性和抑制剂耐药性.
  • 分析不同表达级别的低丰度目标量化和性能.

主要成果:

  • dPCR提供了非常精确的核酸绝对量化,克服了qPCR对标准曲线的需求.
  • 与qPCR不同,dPCR最大限度地减少了与低丰度目标相关的变异性和人工物.
  • 与qPCR相比,dPCR显示出更高的准确性和对抑制剂的抗性.

结论:

  • 对于基因表达分析,dPCR提供了卓越的准确性和稳定性,特别是对于低丰度标.
  • dPCR克服了qPCR的关键局限性,使得结果更加可靠和可重复.
  • dPCR的功能使研究人员能够以 qPCR 不可能的方式研究基因表达.