Microfluidic Whole Genome Amplification Device for Single Cell Sequencing

被引:49
|
作者
Yu, Zhilong [1 ,2 ]
Lu, Sijia [3 ]
Huang, Yanyi [1 ,2 ]
机构
[1] Peking Univ, Biodynam Opt Imaging Ctr BIOPIC, Beijing 100871, Peoples R China
[2] Peking Univ, Coll Engn, Beijing 100871, Peoples R China
[3] Yikon Genom Co Ltd, Taizhou 225300, Jiangsu, Peoples R China
基金
中国国家自然科学基金;
关键词
COPY-NUMBER VARIATIONS; RNA-SEQ; DNA; RECOMBINATION; POLYMERASE; DISEASE; EMBRYOS; PCR;
D O I
10.1021/ac5032176
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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.
引用
收藏
页码:9386 / 9390
页数:5
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