快速高效的方法,并行构建目标的外体和甲基组单链DNA测序库
Eunhye Kim1, Sinae An1,2, Heerak Ahn2
1Department of Pharmacy, School of Pharmacy and Institute of New Drug Development, Jeonbuk National University, Jeonju, 54907, Republic of Korea.
Scientific reports
|February 28, 2025
概括
我们开发了一种高效的工作流程,用于从有限的,受损的DNA中构建并行基因组和表观基因组测序库. 这种方法可以准确地识别儿科脑癌样本中的突变和甲基化,即使是来自老年甲固定组织的样本.
科学领域:
- 基因组学就是基因组学.
- 表观基因组学是指表观基因组学.
- 癌症研究 癌症研究
背景情况:
- 甲素固定和氧化导致的DNA损伤使基因组和表观基因组分析复杂化.
- 很少的DNA输入量限制了测序图书馆的构建效率.
- 儿科大脑癌症通常涉及DNA损伤,需要强大的分析方法.
研究的目的:
- 建立一个高效的工作流程,并行定向的exome和methylome测序库建设.
- 评估该协议的性能损坏的DNA从甲固定嵌 (FFPE) 儿科脑癌样本.
- 评估从低输入,受损DNA获得的测序数据的准确性和统一性.
主要方法:
- 开发一个单链DNA库构建协议.
- 并行向的外体和甲基组测序.
- 适用于9个儿科脑癌FFPEDNA样本的应用.
- 生物信息学对人工突变的过调用.
主要成果:
- 与以前的方法相比,Exome覆盖的统一性更高.
- 89.4-97.0%的体质单核酸变体 (SNV) 的可复制性来自FFPE样本的调用,其DNA输入量为26.7-50ng.
- 获得的甲基化要求78-92%的目标CpG具有100ng的FFPEDNA,与新鲜冷样本相比.
- 从甲基组数据中恢复了39-76%的过SNV.
结论:
- 建立了一个简单,高效的协议,用于并行构建exome和methylome库.
- 该协议对于分析FFPE组织中受损的DNA是有效的,即使存储时间长.
- 这种方法可以对具有挑战性的临床样本进行强大的基因组和表观基因组分析.
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