期刊
ANALYTICAL CHEMISTRY
卷 90, 期 2, 页码 1273-1279出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b04050
关键词
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资金
- National Science Foundation through a CAREER Award [DBI-1253293]
- National Institutes of Health (NIH) [HG007233-01, R01-EB019453-01, 1R21HG007233, DP2-AR068129-01, R01-HG008978, R21AI116218]
- Defense Advanced Research Projects Agency Living Foundries Program [HR0011-12-C-0065, N66001-12-C-4211, HR0011-12-C-0066]
- Fold F(x) Program [DE-AC02-05CH11231]
- NIH [DP2-AR068129-01]
Droplet microfluidics can identify and sort cells using digital reverse transcription polymerase chain reaction (RT-PCR) signals from individual cells. However, current methods require multiple microfabricated devices for enzymatic cell lysis and PCR reagent addition, making the process complex and prone to failure. Here, we describe a new approach that integrates all components into a single device. The method enables controlled exposure of isolated single cells to a high pH buffer, which lyses cells and inactivate's reaction inhibitors but can be instantly neutralized with RT-PCR buffer. Using our chemical lysis approach, we distinguish individual cells' gene expression with data quality equivalent to more complex two-step workflows. Our system accepts cells,and produces droplets ready for amplification, making single-cell droplet RT-PCR faster and more reliable.
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