An addressable dynamic droplet array (aDDA) is presented that combines the advantages of static droplet arrays and continuous-flow droplet platforms. Modular fabrication is employed to create a self-contained integrated aDDA. All the sample preparation steps, including single-cell isolation, cell lysis, amplification, and product retrieval, are performed in sequence within a sub-nanoliter (≈300 pL) droplet. Sequencing-based validation suggests that aDDA reduces the amplification bias of multiple displacement amplification (MDA) and elevates the percentage of one-yeast-cell genome recovery to 91%, as compared to the average of 26% using conventional, 20 µL volume MDA reactions. Thus, aDDA is a valuable addition to the toolbox for high-genome-coverage sequencing of single microbial cells.
Keyphrases
- single cell
- high throughput
- rna seq
- cell cycle arrest
- induced apoptosis
- nucleic acid
- genome wide
- cell death
- breast cancer cells
- pi k akt
- high resolution
- oxidative stress
- high density
- stem cells
- gene expression
- healthcare
- affordable care act
- saccharomyces cerevisiae
- bone marrow
- cell proliferation
- low cost
- tissue engineering
- cell wall