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相关概念视频

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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相关实验视频

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

一种简单,高分辨率的方法来确定DNA结合亲和力和序列选择性.

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
概括
此摘要是机器生成的。

一种新的光间位移 (FID) 测定方法提供了一种简单,快速和具有成本效益的方法来确定DNA结合亲和力和序列选择性. 这种高通量技术分析了与DNA的化合物相互作用,有助于药物发现和开发.

更多相关视频

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

相关实验视频

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

科学领域:

  • 生物化学 生物化学
  • 分子生物学分子生物学
  • 药物发现 药物发现 药物发现

背景情况:

  • 评估DNA结合亲和力和序列选择性对于理解分子相互作用和开发治疗剂至关重要.
  • 现有的方法,如足迹或亲和性切割,可能是耗时的,缺乏高分辨率.
  • 需要一种快速,高吞吐量和成本效益的测试来进行DNA结合分析.

研究的目的:

  • 开发和验证一种简单,非破坏性和高通量方法,用于确定DNA结合亲和性和序列选择性.
  • 为了证明光间位移 (FID) 试验对各种DNA结合剂的分析的实用性.
  • 为了比较定量定位方法来确定DNA结合常数.

主要方法:

  • 开发一种使用乙基化物或提亚色的光间位移 (FID) 测定方法.
  • 采用 96 个井格式的 hairpin deoxyoligonucleotides (512 或 136 个序列) 的库.
  • 通过光损失或内在光变化来评估DNA结合亲和力和序列选择性.

主要成果:

  • FID试验成功生成了化合物的等级级绑定配置文件,以高分辨率定义序列选择性.
  • 这种测试具有成本效益 (约. 100美元/试验),快速 (15分钟读取) 和可自动化.
  • 产生了distamycin A,netropsin,DAPI,Hoechst 33258和berenil的概况,证明了该测试的适用性.

结论:

  • FID测定提供了一种强大,高分辨率的工具,用于表征DNA结合亲和力和序列选择性.
  • 这种方法补充了现有的技术,在速度,成本和吞吐量方面提供了优势.
  • 该测试对药物发现有价值,使得DNA相互作用化合物的有效查和分析成为可能.