Place the phone in the cradle with the camera lens lined up so that it shows a view of the slide from the side (edge-on). Once the camera is set up in the cradle and lined up with the side of the slide, adjust the slide so that the light illuminates the center of the drop and the drop focuses light on the other side of it. After adjusting the slide itself, change the distance of the smartphone from the fluorimeter in order
to keep the drop in focus but keep the view close.
Distance between the smart phone cradle and drop = 5 cm
Solutions Used for Calibration
PCR Product Label Tube
Volume of the DILUTED PCR Product Solution (μL)
Volume of the SYBR GREEN I Dye Solution (μL)
Dilution 1
Dilution 2
Total Dilution (Simplified Fraction)
Positive
80
80
1/6
1/2
1/12
Negative
80
80
1/6
1/2
1/12
G1 1-1
80
80
1/6
1/2
1/12
G1 1-2
80
80
1/6
1/2
1/12
G1 1-3
80
80
1/6
1/2
1/12
G1 2-1
80
80
1/6
1/2
1/12
G1 2-2
80
80
1/6
1/2
1/12
G1 2-3
80
80
1/6
1/2
1/12
Placing Samples onto the Fluorimeter
Find the rough side of the slide (making sure to only touch the slide while wearing gloves)
Place the slide, with the rough side up, in the fluorimeter
Locate the two clear circles in the middle of the slide and add 80 μL of SYBR Green I solution
On the same circle with the SYBR Green I solution, place 80 μL of Sample/Calibration drops
Adjust the slide so that the middle of the drop is illuminated by the light and the drop becomes focused on the side
Data Analysis
Representative Images of Negative and Positive Samples
Image J Values for All Calibrator Samples
Concentration of 2X Calf Thymus DNA solution(micrograms/mL)
Volume of the 2X DNA solution (μL)
Volume of the SYBR GREEN I Dye Solution (μL)
Final DNA concentration in SYBR Green I solution (μg/mL)