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

Figure 2. Digital RT-PCR Taqman assays in microfluidic drops following picoinjection of reverse transcriptase.

Figure 2

(A) Control RT-PCR reactions containing PC3 cell total RNA were emulsified on a T-junction drop maker, thermocycled, and imaged. FAM (green) fluorescence indicates Taqman detection of an EpCAM transcript and Cy5 (red) indicates detection of CD44 transcripts. Brightfield images (BF) of the same drops are shown in the image panel on the far right. The red arrows indicate the direction of emulsion flow in the illustrations. (B) RT-PCR reactions lacking reverse transcriptase were emulsified on a T-junction drop maker and subsequently picoinjected with reverse transcriptase. Picoinjection fluid is pictured as dark gray in the schematic diagram on the left. Brightfield images demonstrate that the drops roughly doubled in size after picoinjection. (C) RT-PCR reactions subjected to picoinjection omitting the reverse transcriptase show no Taqman signal for EpCAM and CD44, demonstrating the specificity of the Taqman assay. Scale bars  = 100 µm.