Virtual microfluidics for digital quantification and single-cell sequencing

被引:0
作者
Xu, Liyi [1 ,2 ]
Brito, Ilana L. [1 ,2 ,3 ]
Alm, Eric J. [1 ,2 ,3 ]
Blainey, Paul C. [1 ,2 ]
机构
[1] MIT, Dept Biol Engn, 77 Massachusetts Ave, Cambridge, MA 02139 USA
[2] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA
[3] MIT, Ctr Microbiome Informat & Therapeut, 77 Massachusetts Ave, Cambridge, MA 02139 USA
关键词
GENOME; AMPLIFICATION; PCR; BACTERIA;
D O I
10.1038/NMETH.3955
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have developed hydrogel-based virtual microfluidics as a simple and robust alternative to complex engineered microfluidic systems for the compartmentalization of nucleic acid amplification reactions. We applied in-gel digital multiple displacement amplification (dMDA) to purified DNA templates, cultured bacterial cells and human microbiome samples in the virtual microfluidics system, and demonstrated whole-genome sequencing of single-cell MDA products with excellent coverage uniformity and markedly reduced chimerism compared with products of liquid MDA reactions.
引用
收藏
页码:759 / 762
页数:4
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