Distance between the smart phone cradle and drop = 4 cm
Solutions Used for Calibration
Concentration ofCalf Thymus DNA solution(micrograms/mL)
Volume of theDNA 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
"[Place the slide, rough side up, onto the fluorimeter]"
"[Place 80 microliters of the SYBR Green Solution into the two middle circles of the slide]"
"[Then place 80 microliters of the calibration solution into the same drop of the SYBR Green solution]"
"[Adjust the slide so the drop is lighted by the blue light of the fluorimeter]"
Data Analysis
Representative Images of Negative and Positive Samples
Positive SampleNegative Sample
Image J Values for All Calibrator Samples
Calibration curve
PCR Results Summary
Our positive control PCR result was 5.44 μg/mL
Our negative control PCR result was -3.19 μg/mL
Observed results
Patient 29148: The first sample was very green
Patient 35264 : The second sample had no color
Conclusions
Patient 29148: This patient was close to the positive control and would amplify nearly as much as the positive control.
Patient 35264 : This patient was close to the negative control and will not amplify as much
SNP Information & Primer Design
Background: About the Disease SNP
SNP is a single nucleotide polymorphism. It occurs commonly within a population in which a single nucleotide (A, T, C, G) differs between members of a biological species. Almost all common SNPS occur within two alleles. SNPs occur within non-coding regions rather than coding regions. Genomic recombination and mutation rate can also vary SNP density. These genetic variations are exploited in DNA fingerprinting which is used in forensic science
Primer Design and Testing
The non-disease primer was successful while the disease primer did not have any results because of the SNPs in the disease primer.