Comparison of multiple displacement amplification (MDA) and multiple annealing and looping-based amplification cycles (MALBAC) in single-cell sequencing.

Comparison of multiple displacement amplification (MDA) and multiple annealing and looping-based amplification cycles (MALBAC) in single-cell sequencing.
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DOI:
10.1371/journal.pone.0114520
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发表时间:
2014
期刊:
影响因子:
3.7
通讯作者:
Ling F
Ling F
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Chen M;Song P;Zou D;Hu X;Zhao S;Gao S;Ling F

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单细胞测序促进了我们对细胞群体异质性的理解,包括不同代细胞之间的单倍型和基因组变异性。全基因组扩增对于为单细胞测序项目产生足够的DNA片段至关重要。利用单精子的测序数据,我们定量比较了两种广泛应用于单细胞测序的主流扩增方法,多重置换扩增(MDA)和多重退火和基于环的扩增循环(MALBAC)。我们的研究结果表明,MALBAC,作为一个修改的MDA和调整PCR的组合,具有更高水平的均匀性,特异性和重复性。
Single-cell sequencing promotes our understanding of the heterogeneity of cellular populations, including the haplotypes and genomic variability among different generation of cells. Whole-genome amplification is crucial to generate sufficient DNA fragments for single-cell sequencing projects. Using sequencing data from single sperms, we quantitatively compare two prevailing amplification methods that extensively applied in single-cell sequencing, multiple displacement amplification (MDA) and multiple annealing and looping-based amplification cycles (MALBAC). Our results show that MALBAC, as a combination of modified MDA and tweaked PCR, has a higher level of uniformity, specificity and reproducibility.
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