Centrifugal step emulsification applied for absolute quantification of nucleic acids by digital droplet RPA
23.04.2015
Schuler F, Schwemmer F, Trotter M, Wadle S, Zengerle R, von Stetten F, Paust Nils
Lab Chip. 2015;15(13):2759-66
We introduce centrifugal step emulsification for the fast and easy production of monodispers droplets. Homogenous droplets with preselectable diameters in a range from 120 ?m to 170 ?m were generated with coefficients of variation of 2-4% and zero run-in time or dead volume. The centrifugal microfluidic droplet generation was used to perform the first digital droplet recombinase polymerase amplification (ddRPA). Compared to digital droplet polymerase chain reaction (ddPCR) the overall processing time of digital analysis was reduced by more than a factor of 4.
