論文

査読有り
2017年12月1日

Massively parallel whole genome amplification for single-cell sequencing using droplet microfluidics

Scientific Reports
  • Masahito Hosokawa
  • ,
  • Yohei Nishikawa
  • ,
  • Masato Kogawa
  • ,
  • Haruko Takeyama

7
1
開始ページ
5199
終了ページ
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1038/s41598-017-05436-4
出版者・発行元
Nature Publishing Group

Massively parallel single-cell genome sequencing is required to further understand genetic diversities in complex biological systems. Whole genome amplification (WGA) is the first step for single-cell sequencing, but its throughput and accuracy are insufficient in conventional reaction platforms. Here, we introduce single droplet multiple displacement amplification (sd-MDA), a method that enables massively parallel amplification of single cell genomes while maintaining sequence accuracy and specificity. Tens of thousands of single cells are compartmentalized in millions of picoliter droplets and then subjected to lysis and WGA by passive droplet fusion in microfluidic channels. Because single cells are isolated in compartments, their genomes are amplified to saturation without contamination. This enables the high-throughput acquisition of contamination-free and cell specific sequence reads from single cells (21,000 single-cells/h), resulting in enhancement of the sequence data quality compared to conventional methods. This method allowed WGA of both single bacterial cells and human cancer cells. The obtained sequencing coverage rivals those of conventional techniques with superior sequence quality. In addition, we also demonstrate de novo assembly of uncultured soil bacteria and obtain draft genomes from single cell sequencing. This sd-MDA is promising for flexible and scalable use in single-cell sequencing.

リンク情報
DOI
https://doi.org/10.1038/s41598-017-05436-4
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/28701744
ID情報
  • DOI : 10.1038/s41598-017-05436-4
  • ISSN : 2045-2322
  • PubMed ID : 28701744
  • SCOPUS ID : 85024126714

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