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相关概念视频

Homologous Recombination02:31

Homologous Recombination

50.2K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
50.2K
Mismatch Repair01:20

Mismatch Repair

4.8K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
4.8K
Labeling DNA Probes03:31

Labeling DNA Probes

8.1K
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...
8.1K
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

11.1K
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...
11.1K
Southern Blot02:57

Southern Blot

18.3K
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...
18.3K

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相关实验视频

Updated: Jun 4, 2025

Author Spotlight: Characterizing Novel Enzymes from Extremophiles and Common Pathogens to Understand DNA Repair and Replication
05:33

Author Spotlight: Characterizing Novel Enzymes from Extremophiles and Common Pathogens to Understand DNA Repair and Replication

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由核酸杂化驱动的酶碎片补充,没有自我标记蛋白质.

Zihan Xu1, Xiaoyu Zhang1, Chandan Pal1

  • 1Department of Chemistry, Binghamton University, The State University of New York, 4400 Vestal Parkway East Binghamton, New York 13902, USA.

Bioorganic chemistry
|December 20, 2024
PubMed
概括

一个新的生物传感器使用酶碎片来检测溶液中的核酸. 这种方法提供了敏感和可编程的DNA和RNA检测,而不需要重的乘客蛋白.

科学领域:

  • 生物化学 生化学
  • 分子生物学分子生物学
  • 生物传感器技术技术

背景情况:

  • 酶片补充试验用于检测生物分子.
  • 目前用于核酸检测的方法可能受到大型组件或序列约束的限制.

研究的目的:

  • 开发和验证用于在稀释水溶液中检测核酸的开启式生物传感器.
  • 创建一个敏感和可编程的检测系统,不需要自标记乘客蛋白质.

主要方法:

  • 使用了NanoBiT的碎片,这是一个分裂的光酶递送酶.
  • 开发的体 (与相关联的寡核酸) 被化为NanoBiT碎片.
  • 使用Drosophila刺自处理域 (DHhC) 进行生物结合.
  • 基于NanoBiT碎片重新关联的示范目标序列激进检测.

主要成果:

  • 在存在低nM度的ssDNA或RNA模板时,实现了NanoBiT发光的30至60倍增加.
  • 开发的生物结合物中没有自我标记的乘客蛋白.
  • 斯特拉默探测器序列显示没有长度或组成的约束.

结论:

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Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
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  • 修改后的NanoBiT-steramer生物结合物作为核酸检测的有效启动生物传感器.
  • 这种方法使得小型DNA和RNA碎片的可编程检测无需序列限制或添加蛋白质批量.
  • 在敏感核酸分析中开辟了新的应用.