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関連する概念動画

DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
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...
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
Modern Molecular Taxonomy01:29

Modern Molecular Taxonomy

Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...

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関連する実験動画

Updated: Jun 18, 2026

DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling
08:04

DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling

Published on: October 8, 2019

DNA結合親和性と配列選択性を確立するためのシンプルで高解像度な方法です.

D L Boger1, B E Fink, S R Brunette

  • 1Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.

Journal of the American Chemical Society
|June 21, 2001
PubMed
まとめ
この要約は機械生成です。

新しい光インターカレーター移位 (FID) 測定法により,DNA結合親和性と配列選択性を決定するためのシンプルで迅速で費用対効果の高い方法が提供されています. この高通量技術では,DNAとの化合物の相互作用をプロファイルし,薬剤の発見と開発を支援します.

さらに関連する動画

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
06:51

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

DNAzyme 10-23 - Based Nanomachines for Nucleic Acid Recognition
07:16

DNAzyme 10-23 - Based Nanomachines for Nucleic Acid Recognition

Published on: February 9, 2024

関連する実験動画

Last Updated: Jun 18, 2026

DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling
08:04

DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling

Published on: October 8, 2019

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage
06:51

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

DNAzyme 10-23 - Based Nanomachines for Nucleic Acid Recognition
07:16

DNAzyme 10-23 - Based Nanomachines for Nucleic Acid Recognition

Published on: February 9, 2024

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.
  • ドラッグ・ディスカバリー・ディスカバリー・ドラッグ・ディスカバリー・ドラッグ・ディスカバリー

背景:

  • DNA結合親和性と配列選択性を評価することは,分子相互作用を理解し,治療薬の開発に不可欠です.
  • フットプリントやアフィニティ・クリバージュなどの既存の方法は,時間がかかり,高解像度が欠けている可能性があります.
  • DNA結合プロファイルのための迅速で,高通量で,費用対効果の高い測定法が必要である.

研究 の 目的:

  • DNA結合親和性と配列選択性を確立するためのシンプルで破壊的でない,高通量メソッドを開発し,検証する.
  • 様々なDNA結合剤のプロファイリングのための光インターカレーター移位 (FID) 解析の有用性を実証する.
  • DNA結合定数を決定するための定量的定位法を比較する.

主な方法:

  • エチジウムブロミドまたはチアゾールオレンジを用いた光インターカレーター移位 (FID) 測定法の開発.
  • ヘアピンデオキシオリゴヌクレオチドのライブラリ (512 または 136 シーケンス) を 96 ウェル形式で使用します.
  • 光喪失または内在光の変化によるDNA結合親和性と配列選択性の評価.

主要な成果:

  • FIDアッセイは,化合物のランクオーダーの結合プロフィールを成功裏に生成し,高解像度で配列選択性を定義しました.
  • この検査は費用対効果が高く (約 $100/アッセイ),高速 (15分読み出し),自動化可能である.
  • ディスタミシンA,ネトロプシン,DAPI,ホーシュト33258,ベリーニルのプロファイルが作成され,アッセイの適用性を実証した.

結論:

  • FIDアッセイは,DNA結合親和性と配列選択性を特徴付けるための強力な高解像度ツールを提供します.
  • この方法は既存のテクニックを補完し,速度,コスト,スループットの点で優位性があります.
  • このアッセイは,薬物の発見に価値があり,DNAと相互作用する化合物の効率的なスクリーニングとプロファイリングを可能にします.