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Comparison of Multiple Displacement Amplification (MDA) and Multiple Annealing and Looping-Based Amplification Cycles (MALBAC) in Limited DNA Sequencing Based on Tube and Droplet.
Micromachines ( IF 3.0 ) Pub Date : 2020-06-29 , DOI: 10.3390/mi11070645
Xiaoxiang Zhou 1 , Ying Xu 1 , Libo Zhu 1 , Zhen Su 1 , Xiaoming Han 1 , Zhen Zhang 1 , Yan Huang 1 , Quanjun Liu 1
Affiliation  

Whole genome amplification (WGA) is crucial for whole genome sequencing to investigate complex genomic alteration at the single-cell or even single-molecule level. Multiple displacement amplification (MDA) and multiple annealing and looping based amplification cycles (MALBAC) are two most widely applied WGA methods, which have different advantages and disadvantages, dependent on research objectives. Herein, we compared the MDA and MALBAC to provide more information on their performance in droplets and tubes. We observed that the droplet method could dramatically reduce the amplification bias and retain the high accuracy of replication than the conventional tube method. Furthermore, the droplet method exhibited higher efficiency and sensitivity for both homozygous and heterozygous single nucleotide variants (SNVs) at the low sequencing depth. In addition, we also found that MALBAC offered a greater uniformity and reproducibility and MDA showed a better efficiency of genomic coverage and SNV detection. Our results provided insights that will allow future decision making.

中文翻译:

基于管和液滴的有限DNA测序中的多个置换扩增(MDA)和多个基于退火和循环的扩增循环(MALBAC)的比较。

全基因组扩增(WGA)对于在单细胞甚至单分子水平上研究复杂的基因组改变对全基因组测序至关重要。多重置换扩增(MDA)和多个基于退火和循环的扩增循环(MALBAC)是两种应用最广泛的WGA方法,取决于研究目的,它们具有不同的优缺点。在这里,我们比较了MDA和MALBAC,以提供有关它们在液滴和试管中的性能的更多信息。我们观察到,与常规试管法相比,微滴法可显着降低扩增偏差并保持高精度的复制。此外,液滴法对低测序深度的纯合和杂合单核苷酸变体(SNV)均显示出更高的效率和灵敏度。此外,我们还发现MALBAC提供了更高的一致性和可重复性,而MDA显示出更高的基因组覆盖率和SNV检测效率。我们的结果提供了可用于将来决策的见解。
更新日期:2020-06-29
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