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関連する概念動画

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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関連する実験動画

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

シングルポリメラーゼ分子からのリアルタイムDNAシーケンシング.

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
まとめ

この研究では,光で標識された核酸およびナノ構造を用いたリアルタイムDNAシーケンシングを実証しています. この方法は高精度で,合成中にポリメラーゼ動態とDNA二次構造を明らかにします.

さらに関連する動画

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

関連する実験動画

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

科学分野:

  • バイオケミストリー バイオケミストリー
  • 分子生物学は分子生物学である.
  • ゲノミクスゲノミクスとは

背景:

  • 精密なDNAシーケンシングは,生物学的プロセスや病気を理解するために不可欠です.
  • 以前の方法は,リアルタイムでの観測と精度の限界に直面していました.

研究 の 目的:

  • 単一分子のリアルタイムDNAシーケンシング方法を開発する.
  • DNAポリメラーゼのダイナミクスを分析し,配列に依存する特性を特定します.

主な方法:

  • 平行単分子検出のためのゼロモード波導体ナノ構造配列を使用しました.
  • 末端リン酸塩と結合した,光で標識されたデオキシリボヌクレオシドトリフォスファート (dNTP) を使用しています.
  • 何千もの塩基を介して,リアルタイムで酵素組み込みをモニターします.

主要な成果:

  • ステリック障害のないDNA合成の継続的な観察を達成しました.
  • DNA二次構造に関連した異なるポリメラーゼ状態と一時停止部位を特定しました.
  • 平均精度99.3%のコンセンサスシーケンスを生成しました.

結論:

  • この新しいシーケンシングアプローチは,ポリメラーゼ運動学への直接的な洞察を提供します.
  • この方法はDNA配列を正確に決定し,ポリメリゼーションの生体物理的パラメータを明らかにします.
  • 高精度と,シーケンス固有のエラーを特定する可能性を証明した.