RT Journal Article
SR Electronic
T1 Oscillating-flow Thermal Gradient PCR
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 544908
DO 10.1101/544908
A1 Varun L. Kopparthy
A1 Niel D. Crews
YR 2019
UL http://biorxiv.org/content/early/2019/02/08/544908.abstract
AB We report the development of a versatile system based on oscillating-flow methodology in a thermal gradient system for nucleic acid analysis. Analysis of DNA and RNA samples were performed in the device, without additional temperature control and complexity. The technique reported in this study eliminates the need for predetermined fluidic channels for thermocycles, and complexity involved with additional incubation steps required for RNA amplification. A microfluidic device was fabricated using rapid prototyping by simply sandwiching dual side adhesive Kapton tape and a PDMS spacer between glass microscope slides. Amplification of the 181-bp segment of a viral phage DNA (ΦX174) and B2M gene in human RNA samples was demonstrated using the system. The developed system enables simultaneous acquisition of amplification and melt curves, eliminating the need for post-processing.