来自美国哈佛大学的谢晓亮教授领导的研究小组开发出一种全基因组扩增方法,称为多重退火和成环循环扩增(MALBAC),使用这种方法对单细胞进行测序,能够获得稳定的高基因组测序覆盖。

-2012年12月21日《科学》

中文翻译


【题目】全基因组测序检测单个人细胞的单核苷酸变异和拷贝数变异

【译文】由于DNA的动态变化,同种细胞可能具有不同的基因组。确定这些基因组差异需要对单细胞进行测序,但是全基因组的扩增偏倚会掩盖这些差异,导致低基因组覆盖。本研究报道了一种新扩增技术——多重退火和成环循环扩增(MALBAC),这项技术能够提供稳定的高基因组测序覆盖。对使用MALBAC技术扩增的DNA进行测序,本研究获得了93%的基因组覆盖,≥1x人单细胞的平均25 x测序结果。本研究检测到单个癌细胞的数字化拷贝数变异(CNVs)。通过对三个同种细胞进行测序,本研究能够鉴定到单个单核苷酸变异(SNVs),且无假阳性结果。本研究直接测定了癌细胞系的全基因组突变频率,同时发现新鉴定的SNVs中嘌呤-嘧啶交换的现象异常频繁。

英文原稿


[Title]: Genome-Wide Detection of Single-Nucleotide and Copy-Number Variations of a Single Human Cell 

[Authors]: Chenghang Zong, Sijia Lu, Alec R. Chapman, and X. Sunney Xie

[Abstract]: Kindred cells can have different genomes because of dynamic changes in DNA. Single-cell sequencing is needed to characterize these genomic differences but has been hindered by whole-genome amplification bias, resulting in low genome coverage. Here, we report on a new amplification method—multiple annealing and looping-based amplification cycles (MALBAC)—that offers high uniformity across the genome. Sequencing MALBAC-amplified DNA achieves 93% genome coverage ≥1x for a single human cell at 25x mean sequencing depth. We detected digitized copy-number variations (CNVs) of a single cancer cell. By sequencing three kindred cells, we were able to identify individual single-nucleotide variations (SNVs), with no false positives detected. We directly measured the genome-wide mutation rate of a cancer cell line and found that purine-pyrimidine exchanges occurred unusually frequently among the newly acquired SNVs.

原文地址

 http://www.sciencemag.org/content/338/6114/1622.abstract

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