Distance between the smart phone cradle and drop = 3 inches
Solutions Used for Calibration
Initial Concentration of 2X Calf Thymus DNA Solution (micrograms/mL)
Volume ofthe 2X DNA Solution (μL)
Volume of the SYBR GREEN I Dye Solution (μL)
Final DNA concentration in SYBR Green I Solution μg/mL
5
80
80
2.5
2
80
80
1
1
80
80
0.5
0.5
80
80
0.25
0.25
80
80
0.125
0
80
80
0
Placing Samples onto the Fluorimeter
Dispense 80μL of SYBR GREEN onto the first two middle dots of the glass tray, make sure the rough end is pointed up.
Dispense 80μL of the Dalf Thymus DNA solution onto the SYBR GREEN drop.
Close the system that the tray is on so that little to no light can penetrate the drop.
Turn on the light and take three bursts of pictures of the drop
Data Analysis
Representative Images of Negative and Positive Samples
Negative:
Positive:
Image J Values for All Calibrator Samples
Final DNA concentration in SYBR Green I solution (µg/mL)
Area
Mean Pixel Value
RAWINTDEN of the drop
RAWINTDEN of the background
RAWINTDEN drop - background
Water
31172
167.519
5221894
2741478
2480416
Water₂
39896
57.972
2312868
154413
2158455
.25 Concentration
36784
200.673
7381553
292217
7089336
.25 Concentration₂
39608
102.807
4071988
168919
3903069
.5 Concentration
31944
208.354
6655650
260365
6395285
.5 Concentration₂
35124
112.853
3963843
142864
3820979
1 Concentration
36500
209.048
7630247
296525
7333722
1 Concentration₂
39588
128.897
5102793
163340
4939453
2 Concentration
37336
236.591
8833359
329838
8503521
2 Concentration₂
40188
194.852
7830707
174987
7655720
5 Concentration
36200
234.436
8486585
291352
8195233
5 Concentration₂
31680
219.348
6948938
153154
6795784
Final DNA concentration in SYBR Green I soluction (µg/mL
RAWINTDEN drop - background
'
'
Standard Deviation
1
2
Mean
Patient 1-1
6530936
2812853
4671894.5
2629081.702
Patient 1-2
5438900
2717659
4078279.5
1924207.964
Patient 1-3
6379485
3602679
4991082
1963498.353
Patient 2-1
6958685
2857649
4908167
2899870.365
Patient 2-2
3976451
8892098
6434274.5
3475887.328
Patient 2-3
9128908
5670777
7399842.5
2445267.88
Calibration curve
PCR Results Summary
Our positive control PCR result was 80 μg/mL
Our negative control PCR result was 80 μg/mL
Observed results
Patient 59797 : The solution was clear, or had a hue of blue due to the flourescent light. This was observed in the three duplicates of the first patient's DNA.
Patient 79049: The solution was clear, or had a hue of blue due to the flourescent light on 2 of the three duplicates. However for the 2nd dupliate, 2-2, the drop was a green color.
Conclusions
Patient 59797 : The patient tested negative for diabetes.
Patient 79049 : The patient testes positive for diabetes.
SNP Information & Primer Design
Background: About the Disease SNP
The SNP was a Pathogenic disease that has an association with coronary heart disease. The variation is found in humans and affects the NHGRI, GWAS, and PheGenl genes.
Primer Design and Testing
For the non-disease DNA code, the result was matched 220bp because there was no mutation on the 3 prime end of the code.
For the disease DNA code, the result was 'no matches' because there was a mutation at the 3 prime end where the A nucleotide aligned to a G nucleotide. Thus causing the mutation and the 'no matches'.