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Updated: May 15, 2025

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在现场进行自动化,高吞吐量的Lytechinus pictus胚胎的杂交

Yoon Lee¹, Chloe Jenniches¹, Rachel Metry¹

¹Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, California, USA.

bioRxiv : the preprint server for biology

|April 8, 2025

概括

我们开发了一个高吞吐量 (HT) -HCR管道,以自动化海胚胎中的基因表达概况,显著提高空间转录学的速度和降低成本.

科学领域:

发展生物学发展生物学
分子生物学分子生物学
基因组学就是基因组学.

背景情况:

传统的现场杂交 (ISH) 方法是劳动密集型的,并且缺乏大规模基因表达分析的吞吐量.
空间转录学对于理解发育过程至关重要,但可扩展的方法有限.

研究的目的:

开发一个全自动化,高通量杂交连锁反应 (HCR) 管道,用于大规模的基因表达造型.
为了降低与海刺, * Lytechinus pictus * 的空间转录组分析相关的成本和手工劳动.

主要方法:

采用小型体积的96井板格式的自动化HCR测定.
整合了通用机器人液体处理器和自动化的共聚焦显微镜.
在32小时内处理了192个基因探针集在整体胚胎上.

主要成果:

为101个目标基因生成了高质量的空间定位数据,这些基因分布在L. pictus*的三个发育阶段.
确定了以前未经表征的生理基因和已知的发育转录因子的局部化模式.
实现了空间转录组数据采集速度的日志顺序增加.

结论:

高吞吐量 (HT) -HCR管道显著提高了海的空间基因表达分析的可扩展性.
这种方法使得研究不足的基因能够有效地进行分析,并促进了复杂的扰动分析.
HT-HCR代表了海发育生物学研究的重大进步.

关键词:

这种植物是Lytechinus pictus.具有高通量功率的高通量杂交连锁反应 (HCR) 是一种在现场混合化 (ISH)在海里,有海,海.空间转录学空间转录学转录因子的转录因子运输商运输商运输商运输商

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相关概念视频

In-situ Hybridization

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...

FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

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,...