植入,决定和放置期间在子宫内组织特异性RNA定位:各种标记方法的技术细微差别
Amanda Dewar1,2, Wenbo Deng1,2,3, Xiaofei Sun1,2
1Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
Current protocols
|June 22, 2023
概括
在现场杂交 (ISH) 在生物样本中定位特定的DNA或RNA序列. 这项研究详细介绍了三种RNA ISH方法,比较了基因表达分析的放射性和非放射性技术.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
- 历史学 历史学 历史学
背景情况:
- 在现场杂交 (ISH) 是一种敏感的技术,用于在细胞和组织中定位特定的DNA或RNA序列.
- 通过使用单链探针来研究空间基因表达模式,RNA ISH是至关重要的.
- 传统的放射性ISH具有局限性,导致开发非放射性方法.
研究的目的:
- 详细介绍和比较三个RNA在位杂交协议.
- 评估放射性与非放射性ISH方法的优缺点.
- 为研究空间基因表达的研究人员提供指南.
主要方法:
- 放射性现场杂交 (放射性-ISH) 是一种放射性现场杂交技术.
- 在位杂交 (DIG-ISH) 的二氧化基因因.
- 滴氧基宁-胺信号放大光素在现场杂交 (DIG-TSA-FISH)
主要成果:
- 非放射性ISH方法提供了传统放射性标签的替代方案,提高了安全性和易用性.
- 泰拉米德信号放大显著增加了检测RNAISH的灵敏度.
- 每种方法在具体性,敏感性和程序复杂性方面都有独特的优缺点.
结论:
- 有各种RNA ISH方法可用于精确的空间基因表达分析.
- 非放射性ISH,特别是带有信号放大,提供了敏感和更安全的替代品.
- 选择合适的ISH协议取决于具体的研究问题和组织类型.
相关概念视频
Regulated mRNA Transport
In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing specific...
In-situ Hybridization
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...


