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相关概念视频

Next-generation Sequencing03:00

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
Sanger Sequencing01:57

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
RNA-seq03:21

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
Lagging Strand Synthesis01:59

Lagging Strand Synthesis

During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
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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相关实验视频

Updated: Jun 27, 2026

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
05:37

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

Published on: April 4, 2025

从单个聚合酶分子进行实时DNA测序.

John Eid1, Adrian Fehr, Jeremy Gray

  • 1Pacific Biosciences, 1505 Adams Drive, Menlo Park, CA 94025, USA.

Science (New York, N.Y.)
|November 22, 2008
PubMed
概括

这项研究展示了使用光标记核酸和纳米结构的实时DNA测序. 该方法实现了高精度,在合成过程中揭示了聚合酶动态和DNA二次结构.

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DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
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Published on: October 6, 2017

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
17:03

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay

Published on: March 23, 2010

相关实验视频

Last Updated: Jun 27, 2026

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
05:37

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

Published on: April 4, 2025

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
07:38

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

Published on: October 6, 2017

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
17:03

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay

Published on: March 23, 2010

科学领域:

  • 生物化学 生物化学
  • 分子生物学分子生物学
  • 基因组学就是基因组学.

背景情况:

  • 精确的DNA测序对于理解生物过程和疾病至关重要.
  • 以前的方法在实时观察和准确性方面遇到了局限性.

研究的目的:

  • 开发一种单分子实时DNA测序方法.
  • 分析DNA聚合酶动态,并确定依赖序列的特征.

主要方法:

  • 利用零模式波导纳米结构阵列进行并行单分子检测.
  • 使用光标记的脱氧核酸三酸盐 (dNTPs) 与终端酸盐结合.
  • 实时监控数千个基的酶合并.

主要成果:

  • 在没有硬质障碍的情况下实现了DNA合成的连续观察.
  • 确定了与DNA二次结构相关的不同聚合酶状态和暂停位.
  • 生成共识序列,平均准确率为99.3%.

结论:

  • 这种新的测序方法为聚合酶动力学提供了直接的见解.
  • 该方法准确地确定DNA序列,并揭示聚合物的生物物理参数.
  • 证明了高精度和识别特定序列错误的潜力.