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PLoS One. 2013 Apr 26;8(4):e62961. doi: 10.1371/journal.pone.0062961. Print 2013.

Picoinjection enables digital detection of RNA with droplet rt-PCR.

Author information

1
Department of Bioengineering and Therapeutic Sciences, California Institute for Quantitative Biosciences, University of California San Francisco, San Francisco, California, USA.

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.

PMID:
23658657
PMCID:
PMC3637249
DOI:
10.1371/journal.pone.0062961
[Indexed for MEDLINE]
Free PMC Article

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