Single-cell genome sequencing at ultra-high-throughput with microfluidic droplet barcoding.
Single-cell genome sequencing at ultra-high-throughput with microfluidic droplet barcoding.
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DOI：
10.1038/nbt.3880
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发表时间：
2017-07
影响因子：
46.9
通讯作者：
Abate AR
中科院分区：
文献类型：
--
作者：
Lan F;Demaree B;Ahmed N;Abate AR
单细胞基因组测序在大细胞群体中的应用受到基因组制备期间分离单细胞的技术挑战的阻碍。在这里，我们提出了单细胞基因组测序（SiC-seq），它使用液滴微流体来分离，片段化和条形码化单细胞的基因组，然后对合并的DNA进行Illumina测序。我们展示了在革兰氏阴性和革兰氏阳性细菌和真菌的合成群落中每次运行> 50，000个细胞的超高通量测序。测序后的基因组可以基于特征序列进行分类。我们使用这种方法来分析分布的抗生素抗性基因，毒力因子，和噬菌体序列的微生物群落从环境样品。常规测序大群体单细胞的能力将使得能够在不同细胞群体中去卷积遗传异质性。
The application of single-cell genome sequencing to large cell populations has been hindered by technical challenges in isolating single cells during genome preparation. Here we present single-cell genomic sequencing (SiC-seq), which uses droplet microfluidics to isolate, fragment, and barcode the genomes of single cells, followed by Illumina sequencing of pooled DNA. We demonstrate ultra-high-throughput sequencing of >50,000 cells per run in a synthetic community of Gram-negative and Gram-positive bacteria and fungi. The sequenced genomes can be sortedin silicobased on characteristic sequences. We use this approach to analyze the distributions of antibiotic-resistance genes, virulence factors, and phage sequences in microbial communities from an environmental sample. The ability to routinely sequence large populations of single cells will enable the de-convolution of genetic heterogeneity in diverse cell populations.
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使用纳米液液滴对单个细胞的高度平行基因组表达分析。
影响因子：
64.5
作者：
Macosko EZ;Basu A;Satija R;Nemesh J;Shekhar K;Goldman M;Tirosh I;Bialas AR;Kamitaki N;Martersteck EM;Trombetta JJ;Weitz DA;Sanes JR;Shalek AK;Regev A;McCarroll SA
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通讯作者：
Mathies, Richard A.
影响因子：
3.7
作者：
de Bourcy CF;De Vlaminck I;Kanbar JN;Wang J;Gawad C;Quake SR
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