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Updated: Jan 29, 2026

Quantitative Real-Time PCR using the Thermo Scientific Solaris qPCR Assay
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Quantitative Real-Time PCR using the Thermo Scientific Solaris qPCR Assay

Published on: June 17, 2010

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实时逆转录定量PCR (RT-qPCR) 方法标准和报告实践

Stephen A Bustin1, Carl T Wittwer2

  • 1Medical Technology Research Centre, Faculty of Health, Education, Medicine and Social Care, Anglia Ruskin University, Chelmsford, United Kingdom.

Clinical chemistry
|January 28, 2026
PubMed
概括

实时逆转录定量PCR (RT-qPCR) 试验的方法报告仍然很差,RNA完整性和PCR效率等关键参数显著下降. 遵守定量PCR实验 (MIQE) 发布最低信息准则是不够的,影响了测试可重复性.

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Real Time RT-PCR02:57

Real Time RT-PCR

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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Transcription01:10

Transcription

Overview
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
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Measurement: Standard Units

Every measurement provides three kinds of information: the size or magnitude of the measurement (a number), a standard of comparison for the measurement (a unit), and an indication of the uncertainty of the measurement. While the number and unit are explicitly represented when a quantity is written, the uncertainty is an aspect of the errors in the measurement results.
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Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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Master Transcription Regulators

Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a  complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
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