Pushbutton-activated microfluidic dropenser for droplet digital PCR

Abstract

Here, we report a portable microfluidic device to generate and dispense droplets simply operated by pushbutton for droplet digital polymerase chain reaction (ddPCR), which is named pushbutton-activated microfluidic dropenser (droplet dispenser) (PAMD). After loading the PCR mixtures and the droplet generation oil to PAMD, digitized PCR mixtures are prepared in PCR tubes after the actuation of a pushbutton. Multiple droplet generation units are simultaneously operated by a single pushbutton, and the size of droplets is controllable by adjusting the geometry of the droplet generation channel. To examine the performance of PAMD, digitized PCR mixtures containing genomic DNA of Escherichia coli (E. coli) O157:H7 prepared by PAMD were assessed by a fluorescence signal analyzer after PCR with a thermal cycler. As a result, PAMD can produce analytical droplets for ddPCR as much as a conventional droplet generator even though any external equipment is not required.