强大的检测基因放大在法甲固定的抛嵌入样本的光 in situ 杂交
Collin Gilbreath1, Yan Peng2, Sihan Wu3
1Children's Medical Center Research Institute, University of Texas Southwestern Medical Center.
Journal of visualized experiments : JoVE
|July 29, 2024
概括
这项研究标准化了光 in situ 杂交 (FISH) 技术,用于检测甲固定嵌 (FFPE) 组织中的基因放大和染色体外DNA (ecDNA). 该协议确保了癌症研究的可复制,高质量的成像.
科学领域:
- 在瘤学瘤学.
- 遗传学 遗传学 是一个
- 分子生物学分子生物学
背景情况:
- 焦点基因放大,包括染色体外DNA (ecDNA),对于癌症的发展和治疗耐药性至关重要.
- 测序方法可以无偏地识别ecDNA,但光现场杂交 (FISH) 对于临床样本来说是成本效益高的.
- 在甲固定嵌 (FFPE) 组织上使用FISH可使基因放大检测,当 kariyotyping 不可能时.
研究的目的:
- 提供一种标准化,优化的协议,用于在FFPE组织样本上使用FISH检测基因放大和ecDNA.
- 解决FFPE组织中FISH分析所面临的挑战.
- 为了实现可复制,高质量的成像,以评估基因放大.
主要方法:
- 开发和优化一个循序渐进的光 in situ 杂交 (FISH) 协议.
- 特别是在甲固化嵌 (FFPE) 组织样本上应用FISH.
- 专注于克服与FFPE样本用于FISH分析相关的独特技术挑战.
主要成果:
- 一个全面的,完全优化的协议,用于FISH检测FFPE组织中的基因放大和ecDNA.
- 标准化程序,使可重现的高质量成像成为可能.
- 在具有挑战性的FPE样本上成功应用FISH.
结论:
- 开发的FISH协议标准化了在FFPE组织中检测基因放大和ecDNA的检测.
- 这种方法为临床试样提供了一种可靠且具有成本效益的方法.
- 研究人员可以获得可复制,高质量的数据来评估癌症研究中的基因放大.
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