当前位置: X-MOL 学术Nat. Biotechnol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Single-cell genome sequencing at ultra-high-throughput with microfluidic droplet barcoding
Nature Biotechnology ( IF 46.9 ) Pub Date : 2017-05-29 00:00:00 , DOI: 10.1038/nbt.3880
Freeman Lan , Benjamin Demaree , Noorsher Ahmed , Adam R Abate

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 sorted in silico based 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.

中文翻译:

超高通量单细胞基因组测序与微流控液滴条形码

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过程中分离单细胞的技术挑战的阻碍。在这里,我们介绍了单细胞基因组测序(SiC-seq),该技术使用液滴微流控技术分离,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社区中,每次运行可> 50,000个细胞的超高通量测序。测序的基因组可以在计算机上分类基于特征序列。我们使用这种方法来分析环境样本中微生物群落中抗生素抗性基因,毒力因子和噬菌体序列的分布。常规地对单个细胞大群体进行测序的能力将使不同细胞群体中遗传异质性的解卷积成为可能。
更新日期:2017-07-13
down
wechat
bug