Distance between the smart phone cradle and drop = 21cm
The cellular device used in the data collecting process requires modifications to the setup. Since the Android device for this collection process does not properly fit the cradle provided, the group balances the Android phone on its bottom and--with the help of a brace--prop up against the elevated fluorimeter. With this setup, it eliminates the issue of stability, is allows easier adjustment if desired , and is easier to deconstruct upon clean-up.
Solutions Used for Calibration
Final DNA concentration in SYBR Green I solution
(The Means Of Each Sample Taken)
Placing Samples onto the Fluorimeter
Step one: Locate and gather all equipment.
Step two: Put on gloves, insert glass (smooth side down) onto fluorimeter
Step three: Calibrate fluorimeter by placing 80μL of SYBR Green I on the first two holes, after adding a new tip, add 80μL of sample onto previous drop
Step four: turn fluorimeter on and adjust slide to align the light beam with the liquid
Step five: Take picture with smartphone. Record distance from phone to liquid
Step six: Turn off fluorimeter, remove slide and clean off liquid
Step seven: Repeat procedures 2-6; substitute 80μL of sample with the subsequent 5 samples
Step eight: Mix unknown PCR from previous week with the red solution provided
Step nine: Proceed through procedures with 80μL SYBR Green I with 5 new mixtures
Data Analysis
Representative Images of Negative and Positive Samples
Image J Values for All Calibrator Samples
Calibration curve
PCR Results Summary
Our positive control PCR result was (on average, if not close to) 0.5 μg/mL
Our negative control PCR result was (on average, if not close to) 0.0 μg/mL
Observed results
Patient #1 : For each sample collect from this patients, the PCR result shows a notable amount of SYBR Green I indicator-coloring present in the solution. Comparing to the results of the calibrations droplets obtained and cataloged (above), it can be concluded this patient possess a significant level of green that suppresses the positive threshold for the disease. Meaning, this patient have contracted the disease and should be quarantined (for safety measures).
Patient #2 : For each sample collect from this patients, the PCR result shows a notable amount of SYBR Green I indicator-coloring present in the solution. However, the amount of the indicator-coloring are significantly less than Patient #1. Comparing to the results of the calibrations droplets obtained and cataloged (above), it can be concluded this patient does not possess a significant level of green that suppresses the positive threshold for the disease; instead, almost meeting the absolute negative threshold. Meaning, this patient does not carries the disease, is clean, and is able to roam around without any worries of infecting others.
Conclusions
SEE ABOVE
SNP Information & Primer Design
Background: About the Disease SNP
SNP is the most common genetic variation. In fact, on average most SNP have little or no negative effect for the human population. Instead SNP is used as a tool for discovery. Importantly, SNP is used by scientists to mark and find the associated diseases. There is much still to discover about SNP. The more we learn, the more beneficial SNP seems to become.
Primer Design and Testing
Using the method of specific primer design can help identify certain portion of A DNA sample after its PCR cycle. Specific primer can be cataloged into databases, with appropriate characteristics associated to a specific sequences. In this case, a reliable online database assists in matching the disease-associated allele found within a series of nucleotides (rs268). From here, the PCR cycle results in the multiplication of this desire portion; assisting in furthering the research of the DNA strand. And since SNP marks the varies degree of differential within a DNA strand, it serves to assist in the crucial discovery of other important differences.
BME 100 Spring 2015
