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多原子单细胞表观基因组剖析的表征和优化.

Leticia Sandoval1,2, Wazim Mohammed Ismail1,2, Amelia Mazzone1,2

  • 1Division of Experimental Pathology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN 55905, USA.

Genes
|June 28, 2023
PubMed
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优化核的隔离对于单细胞多原子试验至关重要. 基于NP-40洗剂的隔离改善了卵巢癌的测序结果,提高了新鲜和冷样本中的细胞类型识别.

科学领域:

  • 单细胞多组的多组学
  • 表观基因组学和转录基因组学.
  • 人类组织分析分析.

背景情况:

  • 像snATAC + snRNA这样的单细胞分析需要高质量的核来准确地表观基因和基因表达概况.
  • 在多原子研究中,优化核隔离方法对于人体组织样本至关重要.
  • 目前的方法需要对可靠性和效率进行评估,特别是对于具有挑战性的样本类型.

研究的目的:

  • 对外围血液单核细胞 (PBMC) 和卵巢癌 (OC) 组织进行不同核隔离技术的比较.
  • 评估隔离方法对测序输出和细胞类型识别的影响.
  • 为了验证原子核隔离对新鲜和冷的人类样本的有效性.

主要方法:

  • 基于NP-40洗剂与基于原酶的核隔离对PBMC和OC的比较.
  • 核体形态和测序数据质量的评估.
  • 评估冷与新鲜样品的准备,以隔离核.
  • 单细胞RNA (scRNA) 和snATAC + snRNA平台的可复制性测试.

主要成果:

  • 与原酶消化相比,基于NP-40洗剂的隔离为卵巢癌提供了优异的测序结果.
  • 细胞核分离方法对细胞类型识别和下游分析产生了重大影响.
关键词:
在表观基因组上,我们进行了表观基因组分析.原子核的制备 原子核的制备一个单细胞测序.这就是 snATAC-seqq.这就是 snRNA-seqq.

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  • 经过适当的准备后,冷和新鲜样品的质量可以比较.
  • 来自scRNA和snRNA测量的基因表达概况证明在细胞识别中具有可比性和有效性.

    • 结论:

    • 选择原子核隔离方法对于成功的单细胞多原子分析至关重要.
    • 建议对卵巢癌组织进行基于NP-40的隔离,以提高数据质量.
    • 在新鲜和冷的人类样本的分析中,snATAC + snRNA平台展示了可重复性和有效性.