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Real Time RT-PCR

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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Updated: Jul 2, 2026

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Published on: November 18, 2014

一种新的miRNA检测方法,使用循环介导的同热放大.

Saiwei Wu¹, Abdu Ahmed Abdullah Al-Maskri², Qun Li¹

¹Department of Pharmacy, The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu, Zhejiang, China.

International journal of analytical chemistry

|September 21, 2023

概括

一种新方法使用基于结合的循环介导同热放大 (miRNA-LAMP) 进行快速和超敏感的微RNA检测. 这种测定为临床诊断和研究提供了敏感的定量分析.

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Published on: November 15, 2017

相关实验视频

Last Updated: Jul 2, 2026

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Published on: November 18, 2014

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Published on: April 14, 2015

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Published on: November 15, 2017

科学领域:

分子生物学分子生物学
生物技术是生物技术.
生物化学生物化学

背景情况:

微RNAs (miRNAs) 是基因表达的关键调节者.
对miRNAs的敏感和快速检测方法是临床诊断和研究所必需的.
对于某些应用,现有的方法可能缺乏所需的灵敏度或速度.

研究的目的:

开发一种新的,快速的,超灵敏的微RNA检测方法.
建立基于结合的循环介导同热放大 (miRNA-LAMP) 系统.
为了能够对特定miRNAs进行高度敏感的定量分析.

主要方法:

设计了两个干环DNA链接探头 (A/B),与目标miRNA杂交.
使用SplintR Ligase进行探针的模板绑定,形成一个子形的amplicon.
采用Bst 2.0 DNA聚合酶和特定的原始剂 (BIP/FIP) 用于循环介导的同热放大 (LAMP).
包含EvaGreen光染料,用于实时检测和量化放大产品.

主要成果:

实现了快速和超敏感的miRNA检测系统.
证明了线性检测范围跨越七个数量级.
为目标miRNA (miR-200a) 确定了100 fM的低检测极限.
通过优化LAMP条件来显著提高放大效率.

结论:

开发的miRNA-LAMP试验具有高度敏感性和定量性.
这种新的方法为精确的miRNA分析提供了新的途径.
该测试为临床诊断和预后研究提供了显著的便利.