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Microfluidic Whole Genome Amplification Device for Single Cell Sequencing

机译:用于单细胞测序的微流体全基因组扩增装置

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We developed a microfluidic device to perform multiplex single-cell whole-genome amplification (WGA) using multiple annealing and looping-based amplification cycles (MALBAC). This device, made of polydimethylsiloxane (PDMS), allows us to monitor the whole process of cell loading and single-cell WGA for sequencing. We show that the genome coverage of MALBAC amplifications is reproducible between chambers on a single chip and between different chips, which enables data normalization using standard samples to accurately identify copy number variations (CNVs). This device provides an easy-to-operate approach to perform single cell sequencing library preparation with minimum hands-on time. It reduces the requirement of manual expertise as well as the risk of contamination, which is essential in future applications especially the medical diagnosis.
机译:我们开发了一种微流体设备,可使用多个退火和基于环的扩增循环(MALBAC)进行多重单细胞全基因组扩增(WGA)。该设备由聚二甲基硅氧烷(PDMS)制成,使我们能够监控细胞加载和单细胞WGA测序的整个过程。我们显示,MALBAC扩增的基因组覆盖范围在单个芯片上的腔室之间以及不同芯片之间是可重现的,这使得使用标准样本进行数据归一化可以准确地识别拷贝数变异(CNV)。该设备提供了一种易于操作的方法,以最少的动手时间即可完成单细胞测序文库的制备。它减少了对人工专业知识的需求以及污染的风险,这对于将来的应用尤其是医疗诊断至关重要。

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