Yu Zhilong, Lu Sijia, Huang Yanyi
Biodynamic Optical Imaging Center (BIOPIC) and College of Engineering, Peking University , Beijing 100871, China.
Anal Chem. 2014 Oct 7;86(19):9386-90. doi: 10.1021/ac5032176. Epub 2014 Sep 22.
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.
我们开发了一种微流控装置,用于使用多次退火和基于环化的扩增循环(MALBAC)进行多重单细胞全基因组扩增(WGA)。该装置由聚二甲基硅氧烷(PDMS)制成,使我们能够监测细胞加载和用于测序的单细胞WGA的全过程。我们表明,MALBAC扩增的基因组覆盖度在单个芯片上的不同腔室之间以及不同芯片之间是可重复的,这使得使用标准样品进行数据归一化以准确识别拷贝数变异(CNV)成为可能。该装置提供了一种易于操作的方法,能够以最少的实际操作时间进行单细胞测序文库制备。它减少了对手动专业技能的要求以及污染风险,这在未来的应用尤其是医学诊断中至关重要。