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Issue 15, 2013
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Bubble-free electrode actuation for micro-preparative scale electrophoresis of RNA

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Abstract

A microfluidic chip is presented for lysis and one-step RNA purification from bacteria. Bacteria are lysed by joule-heating followed by a gel electrophoresis step for clean-up and subsequent elution of small RNA. Bubble formation during electrophoresis at constant current is suppressed through the use of a silver chloride cathode and a silver anode. To prevent silver chloride sediment in the bulk solution, the anode was immersed in a saturated chloride solution. Salt bridges in the form of polyacrylamide gels are used that could be precisely patterned with the help of phaseguides. Bubble-free actuation could be performed for more than 20 min under a constant current. For longer actuation times, cathodic silver-chloride became depleted and a silver-chloride sediment formed in the anodic microchamber at increasing distance from the anode with time. The chip functioning was verified by extraction of transfer-messenger RNA from Escherichia coli and subsequent amplification using reverse transcription real-time PCR. Incorporation of salt bridges enables effective bubble free actuation of Ag/AgCl electrodes in a microfluidic chip. This opens up new possibilities in a surge towards fully integrated diagnostic cartridges that are miniaturized and disposable.

Graphical abstract: Bubble-free electrode actuation for micro-preparative scale electrophoresis of RNA

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Publication details

The article was received on 12 Mar 2013, accepted on 14 May 2013 and first published on 07 Jun 2013


Article type: Paper
DOI: 10.1039/C3LC50332A
Citation: Lab Chip, 2013,13, 2931-2936
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    Bubble-free electrode actuation for micro-preparative scale electrophoresis of RNA

    P. Vulto, P. Kuhn and G. A. Urban, Lab Chip, 2013, 13, 2931
    DOI: 10.1039/C3LC50332A

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