相关实验视频

Updated: Jul 31, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

在单个细胞中通过编码的PCR in situ进行密切位置的单核酸突变的多重化成像

Yao Ren¹, Kerui Liu¹, Hao Yang²

¹College of Biomass Science and Engineering, Healthy Food Evaluation Research Center, Sichuan University, Chengdu 610065, China.

|July 10, 2024

概括

研究人员开发了一种新方法来可视化细菌RNA中的多个密切位置的单核酸突变 (SNMs). 这种技术通过同时成像SNMs来准确地表明病原体的耐药性,从而改善了单个突变检测.

科学领域:

分子生物学分子生物学
微生物学微生物学
遗传学遗传学是一个

背景情况:

在RNA中单核酸突变 (SNM) 与病原体的耐药性有关.
在单细胞中可视化多个密切位置的SNMs对于理解耐药性的异质性至关重要,但仍然具有挑战性.

研究的目的:

开发一种新的方法,以单细胞分辨率在细菌RNA中同时成像多个密切位置的SNMs.
评估该方法在检测*Salmonella enterica*中的诺隆耐药性的有用性.

主要方法:

设计了一种编码的结合介导的 *in situ* 聚合酶链反应 (enPCR) 方法.
使用模块化 anchoring 探头和编码的 PCR 启动器来同时检测 SNM.
将enPCR方法应用于Salmonella enterica*的*gyrA*基因,以检测类抗性突变.

主要成果:

通过enPCR方法,在细菌RNA中成功可视化了多个靠近位置的SNMs.
与单个SNM检测相比,在 *gyrA* 基因中同时检测SNM可以更准确地表明 *S. enterica* 中的类素耐药性.
复合成像试验证明了复杂的基因型和表型之间的关系.

结论:

开发的enPCR方法可以在细菌RNA中可视化多个紧密的SNMs.
这种技术通过同时考虑多个突变,可以更准确地评估病原体的耐药性.
复合成像试验在了解微生物群体中的基因型-表型关系方面具有潜在的应用.

关键词:

抗生素耐药性抗生素耐药性在现场进行PCR.多重复合成像成像技术一个单细胞分析.单核酸突变是一种单核酸突变.

更多相关视频

Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay

Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay

Published on: August 14, 2018

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

Published on: January 7, 2020

相关概念视频

PCR

PCR

Gene Conversion

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

PCR - Polymerase Chain Reaction

PCR - Polymerase Chain Reaction