Picoinjection Enables Digital Detection of RNA with Droplet RT-PCR
Open Access
- 26 April 2013
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 8 (4), e62961
- https://doi.org/10.1371/journal.pone.0062961
Abstract
The ability to add reagents to drops in a sequential fashion is necessary for numerous applications of microfluidics in biology. An important method for accomplishing this is picoinjection, a technique in which reagents are injected into aqueous drops using an electric field. While picoinjection has been shown to allow the precise addition of reagents to drops, its compatibility with biological reactions is yet to be thoroughly demonstrated. Here, we investigate the compatibility of picoinjection with digital RT-PCR Taqman assays, reactions that incorporate nucleic acids, enzymes, and other common biological reagents. We find that picoinjection is compatible with this assay and enables the detection of RNA transcripts at rates comparable to workflows not incorporating picoinjection. We also find that picoinjection results in negligible transfer of material between drops and that the drops faithfully retain their compartmentalization.This publication has 20 references indexed in Scilit:
- High-throughput injection with microfluidics using picoinjectorsProceedings of the National Academy of Sciences of the United States of America, 2010
- Ultrahigh-throughput screening in drop-based microfluidics for directed evolutionProceedings of the National Academy of Sciences of the United States of America, 2010
- Droplet microfluidic technology for single-cell high-throughput screeningProceedings of the National Academy of Sciences of the United States of America, 2009
- Droplet-Based Microfluidic Platforms for the Encapsulation and Screening of Mammalian Cells and Multicellular OrganismsCell Chemical Biology, 2008
- On-Chip Single-Copy Real-Time Reverse-Transcription PCR in Isolated Picoliter DropletsAnalytical Chemistry, 2008
- Reactions in Droplets in Microfluidic ChannelsAngewandte Chemie-International Edition, 2006
- Formation of droplets and bubbles in a microfluidic T-junction—scaling and mechanism of break-upLab on a Chip, 2006
- Droplet fusion by alternating current (AC) field electrocoalescence in microchannelsElectrophoresis, 2005
- Electrocoalescence of water droplets in an insulating liquidJournal of Electrostatics, 1993
- CYTOLOGY OF BURKITT'S TUMOUR (AFRICAN LYMPHOMA)The Lancet, 1964