Single-cell genome sequencing at ultra-high-throughput with microfluidic droplet barcoding.
Freeman LanBenjamin DemareeNoorsher AhmedAdam R AbatePublished in: Nature biotechnology (2017)
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.
Keyphrases
- single cell
- rna seq
- high throughput
- induced apoptosis
- cell cycle arrest
- gram negative
- multidrug resistant
- antibiotic resistance genes
- genome wide
- escherichia coli
- healthcare
- randomized controlled trial
- signaling pathway
- stem cells
- wastewater treatment
- copy number
- clinical trial
- molecular docking
- mental health
- mass spectrometry
- high throughput sequencing
- high resolution
- dna methylation
- mesenchymal stem cells
- anaerobic digestion
- cystic fibrosis
- molecular dynamics simulations
- bone marrow
- molecularly imprinted