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Portable Infrared System for Amplifying DNA and Conducting Field Nucleic Acid TestingPRESENTER: DILLON MIR
Hypothesis
The use of infrared light to heat PCR tubes in order to create a simplified and cost effective method of DNA amplification.
Hypothesis questions
Can we create a model of PCR tube heating using infrared lamps?
Can we create a heating device that can be bought for a low cost or easily made?
Can we make this device easy to use?
Research methods
First tests were run with water in order to create the heating model without using an excess of the somewhat expensive DNA helicase.
Tests were run starting at room temperature (~24°C). Lowered or elevated temperatures causes a difference time to heat the tube.
PCR tube heated to about 65 degrees Celsius.
The heating and cooling models of different temperatures were created in order to find a time and voltage the would not damage the DNA sample.
Allowed to cool to room temperature before another experiment was run.
Heating Equations and derivation
The heat rate accumulation is influenced by the heat flux of the infrared light and the cooling heat flux rate due to convection.
Heating Equations and derivation
Integrating both sides of the equation yields
Solving for the integration constant
Initial Conditions
@t=0 T=To
Cooling Equations and derivation
For periods of cooling, the infrared light is not heating the system anymore.
This reduce the equation just to the cooling rate due to convection.
Cooling Equations and derivation
The value for beta was reduced using the relation between area and volume for a cylinder.
Cooling Equations and derivation
The equation variables are separated and are solved by integrating both sides.
Cooling Equations and derivation
After the integration constant is solved for, it is put back into the equation.
For elevated temperatures
The initial conditions change when working with elevated temperatures.
Initial Conditions
@t=0 T=Te
Prototype 2
The next prototype we created was a “Sun of Vergina” design.
Holds up to 6 PCR tubes for multiple tests at once.
Made of clay with silver paint for reflection.
Future prototype
Created in Solidworks.
Once designed it can be sent to a machine shop to be created.
Easy to produce more with the same specifications.
DNA amplification procedures
DNA amplification process:
1. Combine necessary components together in a PCR tube.
2. Heat PCR tube for 30 minutes.
3. Remove liquid from PCR tube and place in glass slide.
4. turn out light (except blue side LED).
5. Observe drop (green color in drop is amplified DNA).
DNA amplification
Materials Quantity (µL)
Water 14
10X Annealing Buffer 2.5
MgSO4 1
NaCl 2
IsoAmp dNTP Solution 2
DNA template 2.5
Forward Primer 1
Reverse Primer 1
IsoAmp Enzyme Mix 2
DNA amplification
Blue light makes for easier analysis using image software.
Green color inside drop is the amplified (copied) DNA.
Higher concentrations of amplified DNA will produce a more visible shade of green.
Results
The “data” values are the temperature that was measured with respect to time at different voltages.
The “prediction” values are based on the heating model that was created from the collected data.
Conclusions
Using infrared lamps to heat PCR tubes was successful.
Heating and cooling models of a PCR tube with reasonable accuracy were created.
A multi-tube DNA amplification platform was successfully designed and tested.