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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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相关实验视频

Updated: Jul 15, 2025

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

Published on: September 24, 2015

光编码的对数稀释数字滴滴PCR用于超宽动态范围的核酸量化.

Qingyuan Shi¹, Jie Li², Chunchen Liu³

¹State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China; School of Physical Science and Technology, ShanghaiTech University, Shanghai, 201210, China.

Biosensors & bioelectronics

|September 26, 2023

概括

光编码的对数稀释数字滴滴PCR (Flodd-PCR) 显著扩大了核酸检测的动态范围. 这一进步克服了当前数字PCR (dPCR) 方法的局限性,提高了临床适用性.

关键词:

生物感应是一种生物感应.数字PCR是数字的PCR.滴滴滴滴滴滴滴滴滴滴滴滴滴滴滴滴滴滴滴动态范围动态范围这是一个HPVHPVHPV.核酸是核酸中的一种.

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Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR

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Last Updated: Jul 15, 2025

Simple Bulk Readout of Digital Nucleic Acid Quantification Assays

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Published on: September 24, 2015

Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings

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Published on: February 3, 2021

Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR

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Published on: June 26, 2016

科学领域:

分子生物学分子生物学
生物技术是生物技术.
分析化学分析化学

背景情况:

数字PCR (dPCR) 为核酸检测提供了绝对量化和高灵敏度.
与定量PCR (qPCR) 相比,传统的dPCR具有有限的动态范围 (4-5 log),限制了其临床使用.
扩大dPCR的动态范围对于更广泛的应用至关重要.

研究的目的:

开发一种具有扩大动态范围的新型数字PCR方法.
克服传统dPCR的动态范围限制,以提高临床适用性.
以光编码的对数稀释数字滴滴PCR (Flodd-PCR) 呈现为溶液.

主要方法:

使用微流体芯片进行多步序列稀释 (20-925折) 和滴滴生成.
将大约2万个滴滴分为4个组,每个组都有一个独特的DNA模板稀释因子.
采用光编码滴滴用于基于稀释指标的in silico聚类和独立分析.

主要成果:

实现了7级大小的动态范围,超过2级以上的传统dPCR.
已证明可以检测到4-20,000,000副本/μL的合成人类乳头瘤病毒 (HPV) DNA.
在分析临床HPV样本时,Flodd-PCR的性能优于标准dPCR.

结论:

弗洛德PCR显著提高了数字PCR的动态范围,接近qPCR的动态范围.
该方法与现有的dPCR系统兼容,确保了广泛的实用性.
弗洛德-PCR在目前使用传统dPCR的各种应用中显示出前景,特别是在临床诊断中.