Cell2Read:一种自动化工作流程,用于从人类细胞悬浮中生成配序准备的DNA库
Kathryn Whitehead1, Sarah Planchak1, Trinity Williams1
1Center for Biomedical Engineering, School of Engineering, Brown University, Providence, RI 02912, United States.
Biology methods & protocols
|December 30, 2025
概括
Cell2Read自动化了细胞溶解和样本准备用于下一代测序 (NGS),使得从最少1500个细胞中进行高质量的DNA分析. 这种新的方法确保了多种细胞类型的可复制结果,这对于基因组研究和诊断至关重要.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 下一代测序 (NGS) 需要高效的样本准备,特别是从有限的细胞输入.
- 目前的方法可能是劳动密集型的,可能会在细胞数量低或样本类型多样化的情况下扎.
- 需要自动化解决方案来提高基因组分析中的可复制性和可访问性.
研究的目的:
- 引入和验证Cell2Read,这是一个全新的自动化工作流程,用于NGS的集成细胞溶解和样本准备.
- 评估Cell2Read在低细胞输入和各种细胞类型的性能.
- 评估该方法是否适用于分析异质细胞群,包括与瘤学相关的细胞群.
主要方法:
- 开发一个集细胞解离,DNA提取和图书馆准备的自动化平台.
- 优化扩散动力学和热几何学,用于低输入的细胞处理.
- 在多个人类细胞系 (HepG2,Caov3,HEY A8,OVCAR 8,MDA-MB-231) 和初级细胞中进行测试.
- 测序完整性,DNA产量,质量评分,GC偏差和对齐率的评估.
- 评估异质细胞悬浮剂的性能,包括尖癌细胞.
主要成果:
- 在不影响测序完整性的情况下,Cell2Read成功处理了低至1500个细胞的细胞悬浮物.
- 实现了一致的DNA产量 (≥10 ng) 和高的测序质量 (≥3125个细胞的对齐率>95%).
- 该方法在各种细胞类型中证明了可重复性,性能与手动协议相比.
- 实现了异质细胞悬浮的无偏差测序,DNA表示与癌细胞度成比例.
- 该系统从低输入样本中获得了高质量的基因组数据,适合临床诊断和研究.
结论:
- Cell2Read为NGS样本准备提供了一种技术验证,可扩展和自动化的解决方案.
- 该工作流显著扩大了从低投入的人类样本对基因组分析的可访问性.
- 它的坚固性和可重复性使其非常适合瘤学研究和临床诊断.
- Cell2Read确保了高质量,公正的测序数据,支持准确的基因组分析.
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