CIRCULAR WIRE-GUIDED DROPLET THERMOCYCLER FOR RAPID POLYMERASE CHAIN REACTIONS

picture of Roberto Reyes presenting his/her poster: CIRCULAR WIRE-GUIDED DROPLET THERMOCYCLER FOR RAPID POLYMERASE CHAIN REACTIONS

Roberto Reyes , Dustin Karl Harshman, Tu San Park, Jeong-Yeol Yoon

CIRCULAR WIRE-GUIDED DROPLET THERMOCYCLER FOR RAPID POLYMERASE CHAIN REACTIONS

Polymerase chain reaction (PCR) is a very powerful tool in medical diagnostics. For example, PCR is one of only two CDC approved methods of diagnosing H1N1 viral DNA. Conventional PCR typically requires 1-2 hours for completion and reducing the amount of time required for PCR completion is of increasing interest with regards to preventing disease outbreak. In this research, a wire-guided droplet thermocycler was developed in order to achieve faster reaction times. The means involved manipulating a 20 µL droplet through heated silicone oil, in which heat transfer occurs by convection. The thermocycler was designed using AutoCAD software and constructed using a 3D printer. This device consisted of a syringe that manipulated the droplet across three chambers arranged in a circle filled with silicone oil. The oil in each chamber was heated to a different temperature by a circuit heater, controlled by a proportional-integral-derivative algorithm. Droplet insertion and extraction from the syringe was controlled by a motor attached to the syringe plunger. PCR products were verified using gel electrophoresis and UV fluorescent imaging. A control oligonucleotide sequence of ~200 bp was successfully amplified in the presence of a blood sample matrix within 4 minutes 20 seconds. Additionally, a larger bacteria K. pneumonia  was amplified in the same time frame. Further research will focus on finding the detection limit for more realistic samples (i.e. those of lower concentrations and those in other sample matrices such as urine) as well as investigating the properties of droplet PCR (i.e. surface tension, hydrophobicity) that allow for amplification in the presence of blood. The proposed system has potential to become a rapid, bench-top tool for detecting infectious disease. 

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