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Anal Chem. 2017 Oct 3;89(19):10147-10152. doi: 10.1021/acs.analchem.7b02183. Epub 2017 Sep 13.

1D-Reactor Decentralized MDA for Uniform and Accurate Whole Genome Amplification.

Li J1, Lu N1, Shi X1,2,3, Qiao Y1, Chen L1, Duan M1, Hou Y2,3, Ge Q1, Tao Y1, Tu J1, Lu Z1.

Author information

1
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University , Nanjing, 210096, China.
2
BGI-Shenzhen, Shenzhen 518083, China.
3
China National GeneBank, BGI-Shenzhen, Shenzhen 518120, China.

Abstract

Multiple displacement amplification (MDA), a most popular isothermal whole genome amplification (WGA) method, suffers the major hurdle of highly uneven amplification, thus, leading to many problems in approaching biological applications related to copy-number assessment. In addition to the optimization of reagents and conditions, complete physical separation of the entire reaction system into numerous tiny chambers or droplets using microfluidic devices, has been proven efficient to mitigate this amplifying bias in recent works. Here, we present another MDA advance, microchannel MDA (μcMDA), which decentralizes MDA reagents throughout a one-dimensional slender tube. Due to the double effect from soft partition of high molecular-weight DNA molecules and less-limited diffusion of small particles, μcMDA is shown to be significantly effective at improving the amplification uniformity, which enables us to accurately detect single nucleotide variants (SNVs) with higher efficiency and sensitivity. More importantly, this straightforward method requires neither customized instruments nor complicated operations, making it a ready-to-use technique in almost all biological laboratories.

PMID:
28853542
DOI:
10.1021/acs.analchem.7b02183
[Indexed for MEDLINE]

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