Development of a Generic Microfluidic Device for Simultaneous Detection of Antibodies and Nucleic Acids in Oral Fluids
Optimization of the RT-PCR amplicon yield. (a) The effect of different wash buffers on the quality of on-chip RNA isolation was assessed by amplifying increasing amounts of CARD isolated RNA elute by RT-PCR. The volumes represent the amount of eluted RNA used in the amplification reaction using a 10 μL final assay volume. Note the decrease in amplicon yield with increased volume possibly due to the presence of residual EtOH. (b) Doubling of the primers and enzyme concentration and a 2°C lower annealing temperature increased the amplicon yield.