Team:NYU Abu Dhabi/Demonstrate
Demonstrate
In the early stages of experimentation, the team focused mainly on verifying the advantages of RPA compared to other DNA amplification techniques. Throughout extensive comparative research and experimentation, the team optimized the existing RPA protocol using the appropriate volumes for both the reaction mix and sample that yielded better sensitivity and rapidness. Over the course of the optimization process, the team made sure to take into consideration the limitations of the physical diagnostics device that is to embody the aforementioned biological techniques.
Figure 1: Figure 4: SYBR green fluorescence of PCR-amplified HBcAg and ypo2088 samples using bacteria from broth. Left to right: HBcAg 10000 cells/ml, HBcAg 1000 cells/ml, HBcAg 100 cells/ml, ypo2088 10000 cells/ml, ypo2088 1000 cells/ml, ypo2088 100 cells/ml, negative control
In parallel, the engineering team was committed to providing microfluidic cartridges that satisfy all the needs of the wet lab in terms of safety and volumetric precision. The team went through iterations for the microfluidic cartridges using colored TE Buffer to simulate real reagents until there was a clear diffusion area between reagents, no air bubbles were present in channels and the evaporation rate was in an acceptable range.
Figure 2: Fluid flow test on microfluidic chip.
A major feature of the microfluidic cartridges was born out of the need for dispensing small volumes of reagents after a set period of time delay to allow for pre-incubation of preloaded reagents and complete DNA Amplification. The team developed a microfluidic chip capable of both dispensing preloaded fluid into a reaction reservoir, and enable automated actuation. The biology team, being knowledgeable of the design constrictions of the microfluidic chip on which that RPA reaction takes place, selected volumes that can be easily administered into the chip. These volumes were tested using a prototype of the microfluidic chip by running an RPA and verifying DNA amplification through sybr green, nucleic acid stain, fluorescence.
Figure 3: Flourecense test on Volatect's proprietary chip.
