在冷凝固血液样本上的核分离方法
Melissa Cuevas1, Kenneth Jones1, Nancy Hadley Miller1,2
1Department of Orthopedics, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
Bio-protocol
|January 29, 2026
概括
这项研究提出了一种新协议,用于从冷血栓中分离高质量的核,从而使ATAC测序等下游应用成为可能. 这种方法可以保持细胞完整性而不需要冷保存,因此非常适合分析之前收集的血液样本.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 细胞生物学 细胞生物学
背景情况:
- 实验室通常在血清分离后丢弃或不当结血块,从而损害提取的DNA质量.
- 现有的从冷血块中提取DNA的方法可以导致细胞膜脆弱和降低产量.
- 需要一个可靠的协议来从先前冷的血液样本中分离高质量的核材料.
研究的目的:
- 描述一项协议,用于从白细胞中分离高质量的细胞核在以前没有冷保护媒介冷的全血样本中.
- 为了证明这些孤立核的适合性下游基因组应用,包括ATAC测序.
- 通过从冷血栓中分类白细胞的流量进行精确的细胞群分析.
主要方法:
- 在没有冷保护介质的情况下先前冷的全血样本中从白细胞中分离出高质量的核.
- 使用Miltenyi的温和MACS Octo分离器进行样本处理.
- 使用流量分类来表征和分离特定的细胞群,如B细胞和T细胞.
- 通过批量ATAC-seq (对转化酶可访问的染色质序列测试) 验证该协议.
主要成果:
- 成功分离了具有完整核外的核核,适合批量和单细胞测序.
- 证明了孤立核对ATAC测序的有用性,产生了染色质可访问性数据.
- 从先前冷的血栓中特征和分离的B细胞和T细胞.
- 确认了协议与之前冷的血液样本的兼容性,即使没有先前的冷保存.
结论:
- 描述的协议有效地从冷的血块中产生高质量的核,适合高级基因组分析.
- 这种方法克服了与先前冷样本中细胞膜脆弱性相关的局限性.
- 该协议有助于精确的细胞群隔离和下游测序应用,增强档案血液样本的实用性.
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