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从单个细胞开始的exome测序

Ioanna Andreou¹, Markus Storbeck¹, Peter Hahn¹

¹QIAGEN GmbH, Hilden, Germany.

Current protocols

|September 26, 2024

概括

单细胞基因组分析现在可以使用多重位移放大 (MDA) 来进行无偏的DNA放大. 这使得高复杂的下一代测序 (NGS) 库能够在研究中全面检测变异.

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:

单细胞基因组分析提供了对发育,瘤和疾病的见解.
单细胞中的有限DNA传统上挑战了下一代测序 (NGS).
多重位移放大 (MDA) 提供了有限的DNA数量的无偏放大.

研究的目的:

描述一个完整的工作流程,从单个细胞或小量的DNA中生成exome-NGS库.
详细介绍全基因组放大,NGS库生成和外基因组丰富的协议.
促进对起源不明的遗传疾病进行全面的遗传变异分析.

主要方法:

使用多重位移放大 (MDA) 来从单细胞或小数量的DNA中进行无偏向的全基因组放大.
从MDA增强的DNA生成高度复杂的NGS库,以实现高覆盖率.
采用混合捕获丰富方法在预放大后用于外基因组测序.

主要成果:

从单个细胞和少量的DNA中成功生成了exome-NGS库.
NGS库的高收益率和复杂性确保了有效的变体调用.
演示与Illumina测序仪器兼容的完整工作流程.

结论:

MDA技术克服了单细胞基因组分析的DNA限制.
描述的协议使全面的外体序列测序能够识别遗传变异.
这个工作流支持发育生物学,瘤异质性和疾病发病的研究.

关键词:

通过对外组的丰富.基因组学就是基因组学.多重位移放大功率的放大.一个单细胞的单细胞.

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RNA-seq

RNA-seq

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Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...