单分子光现场杂交 (smFISH) 用于细菌中的RNA检测
Camilla Ciolli Mattioli1, Roi Avraham2
1Department of Immunology and Regenerative Biology, Weizmann Institute of Science, Rehovot, Israel. Camilla.ciolli-mattioli@weizmann.ac.il.
Methods in molecular biology (Clifton, N.J.)
|March 19, 2024
概括
本章详细介绍单分子光在位杂交 (smFISH) 用于细菌mRNA量化. 这种可适应的方法提供了对细菌细胞生命的洞察力,并且很容易被其他实验室采用.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 细菌的基因表达调节是复杂的.
- 在细菌中可视化和量化mRNA对于理解细胞过程至关重要.
- 现有的方法可能在适应性或可访问性方面存在局限性.
研究的目的:
- 为在细菌细胞中进行单分子光 in situ 杂交 (smFISH) 提供详细的协议.
- 概述使用smFISH精确量化mRNA转录的方法.
- 突出smFISH在各种细菌研究中的适应性和潜力.
主要方法:
- 在细菌中进行smFISH实验的详细步骤协议.
- 用于成像和分析光标记的mRNA分子的技术.
- 量化每细胞mRNA转录数的策略.
主要成果:
- 成功实施smFISH用于细菌mRNA可视化.
- 在不同细菌物种中展示精确的mRNA量化.
- 在各种实验条件下验证方法的灵活性.
结论:
- 描述的smFISH协议是用于细菌mRNA分析的强大且可适应的技术.
- 这种方法有助于更深入地了解细菌基因表达和细胞动态.
- 该方法易于转移到其他实验室,促进更广泛的采用.
相关概念视频
In-situ Hybridization
9.4K
In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
9.4K
FISH - Fluorescent In-situ Hybridization
20.6K
Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
20.6K
RNA-seq
10.0K
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...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
10.0K


