提取,净化和下一代测序 (NGS) 分析从形式素固定和抛嵌入 (FFPE) 组织中的DNA和RNA
Mainul Hoque1, Veera D'Mello1, Seema Husain1
1Genomics Center, Rutgers New Jersey Medical School, Newark, NJ, USA.
Methods in molecular biology (Clifton, N.J.)
|November 15, 2024
概括
这项研究优化了从甲固定嵌 (FFPE) 组织中提取DNA和RNA的方法. 优化的协议使高质量的DNA测序 (DNA-seq) 和RNA测序 (RNA-seq) 图书馆为FFPE样本做好准备.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 组织病理学 组织病理学
背景情况:
- 嵌入式固定 (FFPE) 组织是有价值的生物资源,因为它们在室温下具有长期存储稳定性.
- 形式素固定诱导核酸中的化学修饰和降解,对分子分析构成挑战.
- 尽管面临挑战,但过去20年的进步提高了FFPE组织对DNA和RNA分析的实用性,包括下一代测序.
研究的目的:
- 描述从FFPE组织中顺序提取DNA和RNA的优化方法.
- 详细说明从FFPE衍生核酸中制备DNA测序 (DNA-seq) 和RNA测序 (RNA-seq) 库的准备.
- 为了促进FFPE样本在Illumina平台上的下游基因组应用中使用.
主要方法:
- 从FFPE组织切片中顺序提取DNA和RNA.
- 提取协议的优化,以最大限度地提高产量并最大限度地降低退化.
- 使用商用套件准备DNA-seq和RNA-seq库.
- 应用Illumina测序平台用于图书馆分析.
主要成果:
- 从FFPE组织中证明了DNA和RNA的顺序提取成功.
- 获得了高质量的核酸产量,适合用于图书馆准备.
- 生成了强大的DNA-seq和RNA-seq库,与Illumina测序兼容.
- 验证了FFPE样本可靠基因组分析的优化方法.
结论:
- 优化的序列提取和库准备方法提高了FFPE组织对综合基因组研究的实用性.
- 这些方法为从档案FFPE样本中进行DNA和RNA分析提供了可靠的方法.
- 描述的协议支持将下一代测序技术应用于FPPE衍生的核酸,扩大研究可能性.
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